Mitochondria

Mitochondria were first observed by Altman (1894) as filamentous structures and he

called them as bioblasts, but Brenda (1897) called them as mitochondria. Unlike
chloroplasts they are found in almost all eukaryo c organisms. They are very important
energy rich ATP producing organelles and make it available for other biological processes
in the form of energy rich compounds like ATP and NADH2 and NADPH2. It also directs
fa y acid oxida on products for ATP synthesis. As an endosymbiont mitochondria
originated from bacterial cells approximately 1.2 to 2 billion years ago. Origin by a
process called symbiogenesis ar culated by a Russian Botanist Konstantin Mereschowsky
in 1910. A recent study by researchers of the University of Hawaii at Manoa and
the Oregon State University indicates that the SAR11 clade of bacteria shares a rela vely
recent common ancestor with the mitochondria exis ng in most eukaryo c cells.
Cyanobacteria and α-proteobacteria are the most closely related free-living organisms
to plas ds and mitochondria respec vely. All eukaryote contain mitochondria in their
cells, except red blood cell. With endosymbiosis the bacterial cells lost their cell wall,
but added one more membrane and lost most of its DNA to host eukaryo c cell, but
remained together coopera vely. At least in human popula on both males and females
have mitochondria, but inherited to offspring’s only from females.

http://biogenevent.weebly.com /h p://bio100.class.uic.edu/
h p://learn.gene cs.utah.edu/content/begin/cells/organelles/

https://universe-ureview.caUniverse https://

h p://unicerse-review.ca

Analysis of mitochondrial DNA from people around the world has revealed many clues about ancient
human migra on pa erns. h p://learn.gene cs.utah.edu/

Schema c representa on, as shown in the above diagrams, of endosymbiosis, called

‘symbiosgenesis’ leading to the appearance of mitochondria; then there is succession of
endosymbiosis with cyanobacteria, what will lead to the crea on of a second new
organelle specific for plant cell; Chloroplast.; the uniqueness of both mitochondria and
chloroplast is that they replicate independently from nuclear DNA replica on- for they
have their own DNA and their own mechanism of replica on, however they require
nuclear inputs also for in the past when they are taken in they have lost most of their
DNA to the nucleus, but they have coordination with each other. It is estimated that
primary plastid s in eukaryotes have believed to have occurred approximately 1.5 billion
years ago. It is controversial indeed. Chloroplast endosymbiosis occurred after
mitochondria.
The idea of endosymbiosis was first proposed by Konstan n Mereschkowski, a prominent Russian
biologist, in 1905. He coined the term "symbiogenesis" when he observed the symbio c
rela onship between fungi and algae.
The term "endosymbiosis" has a Greek origin (endo, meaning "within"; syn, meaning "with";
and biosis, meaning "living"), and it refers to the phenomenon of an organism living within
another organism. In 1923, American biologist Ivan Wallin expanded on this theory when he
explained the origin of mitochondria in eukaryotes (Wallin 1923). However, not un l the 1960s
did Lynn Margulis, as a young faculty member at Boston University, substan ate the
endosymbio c hypothesis. Based on cytological, biochemical, and paleontological evidence, she
proposed that endosymbiosis was the means by which mitochondria and plas ds originated in
eukaryotes (Sagan 1967; Margulis 1970). In those days, the research community viewed her
unconven onal idea with much skep cism, but her work was eventually published in 1967
(Sagan 1967) a er being rejected by fi een scien fic journals! Today, endosymbiosis is a widely
accepted hypothesis to explain the origin of intracellular organelles. h p://www.nature.com/

One model for the origin of mitochondria and plastids. https://en.wikipedia.org/

h p://www.enchantedlearning.com/galleryhip.comwww.cas.miamioh.edu

Mitochondria in Sieve cells (M) of Vicia faba; sieve tube cell cross wall is perforated through which
Plasmodesmata pass though which provides symplastic connection;. One finds protoplasmic layers next to
Plamamembrane, companion cells are adjacent and tightly adhered and one finds a symplastic and as well
as aplasic liquid movement, also one finds protein bodies called Pi, endoplasmic reticulum is visible and
even mitochondria are found; plastids are also found, Apparently no nucleus is found. In some case certain
sieve tube cell associated parenchyma cells are transformed into Transfer cells; http://public.wsu.edu/

Schema c drawing of sieve tube structure in Arabidopsis thaliana; C = chloroplast, Cl = clamp proteins, ER
= endoplasmic re culum, EV = electron dense vesicles, GM = ground matrix, M = mitochondrium, N =
nucleus, P = plas d, SR = SEOR1 filaments, V = vacuole. From Froelich et al. 2011. Phloem ultrastructure
and pressure flow: SEOR protein agglomera ons do not affect transloca on. Plant Cell . Plant
Cell doi/10.1105/tpc.111.093179 Copyright American Society of Plant Biologists; h p://public.wsu.edu/

https://mortada8.wordpress.com/2009

h p://faculty.fmcc.suny.edu/

www.biologyonline.us

Mitochondria are the cells’ microscopic power plants, producing ATP which powers your
muscles; h ps://roboplant.wordpress.com

EM of mitochondria- showing inner membrane bond Elementary par cles involved in ATP produc on;
h p://lifesci.dls.rutgers.edu/

Elementary par cles (ATP synthases); h p://biology4isc.weebly.com/

Mitochondrial DNA circular; www.visualphotos.com

Shape sand Size:

Using specific stains or fluorescent dyes, mitochondia can be observed cytologically. In
the presence of janus green they look like small rod shaped, greenish blue structures.
The shape of mitochondria is never constant; at one moment they look like small
bacterial shaped or tubular shaped structures of 0-.5 mm to 7 mm and in next few
moments, they appear as long filaments or vesicles. A movie on mitochondria, pictured
to show its variability in structure with me, it is amazing to observe how the
mitochondria divide and fuse with one another and divide - a process con nuum,
perhaps in all organisms. Thus they exhibit polymorphism in their shape and size.

Another, more recent discovery about the structure of mitochondria is the close associa on that they can
have with the endoplasmic re culum (ER), called the mitochondria-associated ER membrane or MAM.
Mitochondria store signaling molecules like Ca2+; Mitochondria can also par cipate in cell signaling
through the reac ve oxygen species (ROS) that are created during
metabolism. h p://frombenchtobedside.wordpress.com/

Communication with each other in terms of chemicals they exchange: www.glogster.com

Ivan Jaijic et al;h p://www.mdpi.com/

Senescence, Stress, and Reactive Oxygen Species:

Generation of reactive oxygen species (ROS) is one of the earliest responses of plant cells to
various biotic and abiotic stresses. ROS are capable of inducing cellular damage by oxidation of
proteins, inactivation of enzymes, alterations in the gene expression, and decomposition of
biomembranes. On the other hand, they also have a signaling role and changes in production of
ROS can act as signals that change the transcription of genes that favor the acclimation of plants
to abiotic stresses. Among the ROS, it is believed that H2O2causes the largest changes in the
levels of gene expression in plants. A wide range of plant responses has been found to be
triggered by H2O2 such as acclimation to drought, photooxidative stress, and induction of
senescence. Our knowledge on signaling roles of singlet oxygen (1O2) has been limited by its
short lifetime, but recent experiments with a flumutant demonstrated that singlet oxygen does not
act primarily as a toxin but rather as a signal that activates several stress-response pathways. In
this review we summarize the latest progress on the signaling roles of ROS during senescence
and abiotic stresses and we give a short overview of the methods that can be used for their
assessment. Ivan Jaijic et al;h p://www.mdpi.com/

Schematic model of ascorbate transport in chloroplasts.

Upon photostress, PHT4;4 gene expression is enhanced, and thePHT4;4protein at the envelope
membranes takes up ascorbate from mitochondria, which is transferred into the thylakoid through
an as yet unknown transporter. PHT4;1 is a candidate ascorbate transporter at the thylakoid
membrane; Takaaki Miyaji et al. http://www.nature.com/

Mitochondria are involved in regulating the cell cycle and play a key role in the apoptosis
pathway, a type of programmed cell death. And just for the sake of completeness: some parts of
the steroid hormone, heme synthesis and urea cycle pathways take place in mitochondria.
http://frombenchtobedside.wordpress.com/

Number and Distribu on:

The number of mitochondria found in cells varies from cell type to cell type; it varies
from species to species. Even physiological state of the cell determines the number.
Trypanosome cells contain only one mitochondrium. Yeast cells under glucose
repression posses just one or two mitochondria per cell. But a liver or meristems in
contain 500-1600 mitochondria per cell. In some amphibian oocytes which are very
ac ve, there can be 10,0000 or more mitochondria. The cell that requires more energy
contains more number of mitochondria, ex: flight muscle cells in insects, heart cells have
more mitochondria than their intes nal cells.
Even the intracellular distribu on and orienta on of mitochondria depends upon the
structures involved in a par culars func on. For example, in flight muscle cells, in the
vicinity of centrosomes, mitochondria are arranged radially and mitochondria are
packed longitudinally. At the basal granules of flagella they are aggregated at the base;
otherwise they are randomly distributed in the cytoplasm. Mitochondrial dysfunc on
can cause aging, and found to operate in diabetes 2 and Alzheimer’s disease

Structure:
Probably mitochondrion is the only cell organelle whose structure has been studied in
greater details. Like any other cell organelles, it is also bounded by membranes. The
outer membrane is 6 nm thick and the length varies from mitochondria to mitochondria
and from to cell. Structurally mitochondrion consists of two membranes, the outer and
the inner membranes and they are separated from each other with perimitochondrial
space. The outer membrane can be striped of from the inner membrane by dgitonin
treatment and such mitochondria are called mitoplasts.
The outer membrane can be easily iden fied by its marker enzyme called monoamine
oxidase. The outer surface of the outer membrane appears to be smooth but careful
observa ons reveal the presence of many aggregated granular structures which are
believed to be enzymes responsible for glycoly c process. The perimitochondrial space
is filled with a fluid in which, enzymes like Adenosyl cyclases Cyt.c and others are found.
The inner membrane shows structural complexity which is unique to this organelle. It is
folded into a number of membranous crests called cristae. They may be oriented
transversely or longitudinally. But the number of cristae varies and it depends upon the
func onal state of the cell. For example, mitochondria found in flight muscle cells of
insects contain 10,000 or more cristae, but in res ng cells the number of cristae in
mitochondria is few. The increase in the number of cristae increases the surface area of
reac on which helps in the produc on of more of ATPs.
Though cristae provide a kind of compartmenta on to mitochondrial chamber, it is not
total. The perimitochondrial space is con nuous in the cristae folds. In spite of both
membranes being dis nct, they are in contact with each other at least at 70-100 sites.
Recent studies, involving mitochondrial contrac on and expansion due to changes in
energy levels of mitochondria, have demonstrated that most of the mitochondria
exhibit several contact zones with ER; where at the cytoplasmic surface a cluster of
ribosomal complexes are found. It is speculated that it is through this region that some
of polypep des synthesized in the cytosol are transported into mitochondia. Probably
these regions are specialized structures involved in the transporta on of components
from cytoplasm into mitochondria. Now it is known that there are specific transporters
of nuclear coded proteins are transported into mitochondria through such transporters.
The outer membrane has Tom and the inner membrane has Tim. These have been
characterized. Basically these contain a receptor protein and channel proteins.
The inner surface of the cristae membranes ie, towards mitochondrial matrix side, is
studded with a large number of knob or globular shaped par cles with a conspicuous
stalk like structure. Such globular par culate are called Racker par cles which have
been iden fied as F1 par cles. They act as ATP synthetases and they are uniformly
spaced at 10 nm apart. The number of F1 par cles present in each mitochondria ranges
from 100 to 1000. The cristae membranes also contain a large number of electron
transpor ng enzymes and other oxido reductases deeply buried in the core of the lipid
bilayers, but vectorially organized. In fact, electron transpor ng enzyma c units
cons tute more than 20% of the total membrane proteins.
Mitochondrial chamber is filled with fluid called mitochondrial matrix. A host of
enzymes are present in this fluid. The unique feature of matrix is the presence of a
circular DNA duplex and all t RNAs, and mRNAs. It also contains 70s ribosomes which
are similar to that of prokaryotes. Having the above said structures, mitochondrial is
considered as a semiautonomous organelle.

Chemical Composi on and Func ons:

The outer membrane consists of 40% lipids and 60 percent proteins. The common
liquids found are lecithin, cephalin and cholesterol. Among proteins, along with
structural ones, the outer membrane posses a marker enzyme called monoamine
oxidase. On the contrary, the inner membrane is made up of 20% lipids and 80%
proteins. The notable lipids present are cholesterol, lecithin and cephalin and
cardiolipins, the la er is present is significant amount. Among proteins present in the
inner membrane are ATP synthetase succinic dehydrogenase carrier proteins like
carni ne, fa y acid acyl transferase, ATP and ADP carrier, which transport inorganic
phosphates, calcium, malate, glutamate, and many others.
The electron transport enzymes, proton secre ng proteins are virtually buried in the
core of the inner membranes. However, electron transpor ng proteins, which include
oxido-reductases, are grouped into four complexes. Each of them vectorially arranged
for receiving the reduced coenzymes like FADH2 and NADH2 which are then subjected
to terminal oxida on, while the outer is virtually impermeable even for simple ions.
They have to be transported across the membrane through specific carriers.
Mitochondrial matrix also consists of a wide variety of enzymes for kreb’s cycle, amino
acid metabolism, protein metabolism, nucleic acid metabolism and fa y acid oxida on.
Mitochondrial DNA found in the matrix is found in mul ple copies from 40 to 60 or
more. The DNA is in circular form and codes for 2rRNAs, all 22tRNAs and 13
mitochondrial proteins. Mammalian mitochondria of human contains a circular DNA of
~16500 bases pairs, but yeast mit is 2-3 mes larger than mammalian and it has been
mapped. Among 13 proteins coding genes, three for 3 large polypep des of
cytochrome oxidase, and one for the large subunit of cyt.B-c reductase and one
hydrophobic protein “transfer protein” of ATPase complex are present. The other gene
products which have not been iden fied are called URF1, URF2 etc. The unique feature
of mit DNA is that most of the gene products are transcribed on H strand, but L strand in
also used for the produc on of transcripts for few tRNAs and one or two URFs. In
trypanosome only one mitochondrium is present and it contains Kinetoplas da. It has a
large single maxi chromosomal DNA and hundreds of smaller min circular DNAs.
The DNA replica on in mitochondria is independent or some me synchronized with
that of nuclear DNA replica on. But the mechanism of replica on, though it is
semiconserva ve it exhibits D-loop mechanism. Another important feature of mit DNA
is all genes are ghtly packed with small spacers or intervening sequences in between
the genes or within the transcribing genes. tRNA genes are located in between protein
coding frames. However, yeast mitochondria have a long DNA with redundant and
intervening sequences within the coding sequences. In plants, mitochondrial DNAs,
besides having the above said genes they also have a gene for male sterility, ex: Zea
mays.

Mitochondrial Transla onal Machinery:

The ribosomal machinery present in mitochondrial matrix is typical of prokaryo c 70s
type, but in some plants the ribosomes presents in mitochondria are slightly smaller
than 70s. Nevertheless the 23s RNA, 16s rRNA and 5s RNA are present with some minor
differences. Most of the transcribing the transla onal factors and other enzymes found
in mitochondrial matrix are coded for by nuclear genome and translated on cytosol 80s
ribosomes, then they are imported into mitochondria. From the ribosomes it is clear
that mitochondrial matrix has all the components for nucleic acid & protein synthesis.
Mitochondrial matrix is also involved in citric acid cycle, where energy rich co-enzymes
like FADH2 and NADH2 are produced. They are then taken up by cristae membranes,
where they are step wisely and sequen ally oxidized. During this process, the energy
released is used for the synthesis of ATP. This phenomenon is called oxida ve
phosphoryla on. These mitochondria are endowed with a unique structural and
func onal co-ordina on.
Mitochondrial Genome:

Organization of the mammalian mitochondrial genome. Thirteen protein-coding genes

(yellow), twenty-two tRNA genes (red) and two rRNA genes (orange) are encoded on a single
circular nucleic acid and transcribed from three promoters (blue): LSP, HSP1 and HSP2, which
are situated in a single region called the D-loop, which contains regulatory sequences that
control transcription from all three promoters, including motifs for DNA-binding proteins such
as Tfam. The inner circle of genes is encoded on the (-) strand and transcribed from the LSP
promoter. The outer circle of genes is encoded on the (+) strand and transcribed from the
HSP1 and HSP2 promoters. Transcription from HSP2 is terminated distal to the 16S rRNA
gene. The resulting three polycistronic transcripts are processed by enzymatic excision of the
tRNAs (red). ATP6, ATP8, subunits of ATP synthase F0; Cox1, Cox2, Cox3, subunits of
cytochrome oxidase; CytB, cytochrome B, Nd1, Nd2, Nd3, Nd4, Nd4L, Nd5, Nd6, subunits of
NADH dehydrogenase. http://www.genomebiology.com/

Heidi Chial, Ph.D. ;www.nature.com

Human Mit DNA is circular and 16,569bp long; Mit DNA contains 37 genes, that
codes for 13 proteins, 22 tRNAs and 2 rRNAs; The mitochondrial genome is circular,
whereas the nuclear genome is linear (Figure above).
The mitochondrial genome is built of 16,569 DNA base pairs, whereas the nuclear
genome is made of ~3.3 billion DNA base pairs.
The mitochondrial genome contains 37 genes that encode 13 proteins, 22 tRNAs,
and 2 rRNAs.
The 13 mitochondrial gene-encoded proteins all instruct cells to produce protein
subunits of the enzyme complexes of the oxida ve phosphoryla on system, which
enables mitochondria to act as the powerhouses of our cells.
The small mitochondrial genome is not able to independently produce all of the
proteins needed for func onality; thus, mitochondria rely heavily on imported
nuclear gene products.
One mitochondrion contains dozens of copies of its mitochondrial genome. In
addi on, each cell contains numerous mitochondria. Therefore, a given cell can contain
several thousand copies of its mitochondrial genome, but only one copy of its nuclear
genome.

The mitochondrial genome is not enveloped, and is it not packaged

into chroma n.
The mitochondrial genome contains few, if any, noncoding DNA sequences. (Three
percent of the mitochondrial genome is noncoding DNA, whereas 93% of the nuclear
genome is noncoding DNA).
Some mitochondrial coding sequences (triplet codons) do not follow
the universal codon usage rules when they are translated into proteins.
Some mitochondrial nucleo de bases exhibit func onal overlap between two
genes; in other words, the same nucleo de can some mes func on as both the last
base of one gene and the first base of the next gene.
The mitochondrial mode of inheritance is strictly maternal, whereas nuclear
genomes are inherited equally from both parents. Therefore, mitochondria-associated
disease muta ons are also always inherited maternally.
Mitochondrial DNA is coded by both strands, for they transcribe and generate
rRNA, tRNA and mRNAs.

Mitochondrial genes on both DNA strands are transcribed in a polycistronic manner:

Large mitochondrial mRNAs contain the instruc ons to build many different proteins,
which are encoded one a er the next along the mRNA. In contrast, nuclear genes are
usually transcribed one at a me from their own mRNA. h p://www.nature.com/
Mitochondrial muta ons can lead to aging and cancer. Mitochondrial disease are many
such as Leber hereditary op c neuropathy (LHON), causes postlingual deafness
(deafness that occurs a er three years of age, when a child has already learned to
speak), diabetes, examples include Pearson syndrome, Leigh syndrome, progressive
external ophthalmoplegia, exercise-induced muscle pain, fa gue, and rhabdomyolysis
and many others.

Biogenesis:
In the past 20-30 years various specula ve theories have been proposed from me to
me to explain mitochondrial biogenesis. Most of the views were based on cytological
observa ons. But now the structure, gene c composi on and its func onal aspects are
fairly known. Recent studies clearly indicate that mitochondria originate from the pre
exis ng mitochondria by growth and fission. The biogenesis of mitochondria is highly
regulated and co-ordinates with the nuclear genome. Though, more than 95% of the
total mitochondrial proteins are coded for by nuclear genome but certain factors coded
for by the nuclear genome exert control over mitochondria gene expression.

Coordina on of plant mitochondrial biogenesis: keeping pace with cellular requirements ;Elina Welchen
etal, h p://journal.fron ersin.org/

Similarly, some mitochondrial protein factors coded for by the mitochondrial genes are
transported into nucleus where they regulate the expression of genes and gene
products required for mitochondria (one report from Neurospora). In this coordinated
network, mitochondria need to generate signals to modify nuclear gene expression
according to organelle and cellular requirements. Signaling pathways from organelles to
the nucleus are referred to as Retrograde Signaling pathways. While several relevant
players in chloroplast retrograde signaling have been extensively characterized (for
reviews see Leister et al., 2011;Kleine and Leister, 2013), rela vely li le is known about
mitochondrial retrograde signaling (MRS). Considering that chloroplasts and
mitochondria are closely connected, signaling pathways involved in chloroplast
retrograde regula on need to be explored in the context of mitochondria to get new
insights into MRS.

PGC1-alpha mediated pathway governing mitochondrial biogenesis and func on; certain transcrip on
factors NRF-1, NRF-2, ERRa, PPARa and MEF-2 act on PGC-1aloha; h p://physrev.physiology.org/

As mitochondrial genome and 70s ribosomal machinery and other transcrip on

transla ng factors are more or less similar to that of prokaryo c bacteria, it is viewed by
many biologists that the eukaryo c mitochondria were once bacterial cells; in course of
me they have incorporated into eukaryo c cells properly for ~3 billion years ago. Since
then they exist as symbionts. The similari es between bacteria and mitochondria are
circular DNA, chloramphenicol sensi ve,70s ribosomes, 23s RNA, 16s RNA and 5s RNA.
Based on these observa ons, it is now strongly believed that mitochondria are symbio c
bacteria; now it is an accepted fact. Mitochondrial codons are not universal.
Mitochondrial DNA has been used to find out phylogeny. It is now believed all the
present 6 billion humans have derived mitochondria from few African mothers.
Mitochondrial DNA gives foot print of human race; for example 4.5000 to 5000 years
ago human popula on lived around Indus valley, are now called with the arrival of
Aryans (now called Brahmins, most of them from Iran called Hindus). Today take all
casts and creeds of human popula on in India, if one observes mit. DNA one finds this
Indus valley lived women for only women contribute mitochondria to their offspring’s.
The origin of humans took place in Africa 120,000-165000 years ago, this popula on
moved to India via Indus valley around 60,000 years ago, and this popula on moved to
China and Australia 50k and Europe around 40K ago. Indian women have its imprint in
the form of Mitochondrial DNA, which provided inheritance pa ern that we see in
human popula on whatever cast or communi es you belong. You cannot ques on the
poof of DNA. You cannot ques on this gene c inheritance, so also Male Y chromosomal
DNA.

Prof. Aleksandra Trifunovic; www.cecad.uni-koeln.de

Kinetoplas da; h ps://en.wikipedia.org; h p://www.nature.com/

Kinetoplas d DNA (kDNA) is organized into an incredible network of interlocked rings.

The kDNA codes for cytochrome b oxidase subunits, NADH dehydrogenase subunits,
ATPase subunit 6 and rRNAs By studying this amazing structure, how did scien sts learn
about RNA edi ng?
Laura Vargas-Parada,2010 nature.; www.nature.com

Concatenated DNA of the Kinetoplast of kinetoplastids (Trypanosoma species and

Leishmania species etc).
Very interes ng is the kinetoplas d, this structure is present in Trypanosoma cruzi, T.
rhodesience, T.gambiense, T.brucei and other similar trypanosomes.. It contains a single
mitochondrium and dense granule within its mitochondria. This structure consists of
concatenated circular DNA and they are associated with structural proteins and RNA
polymerases. This kinetoplas d is found at the base of the flagella. The DNA is very
unusual for it has two different DNA, one maxicircle and another minicircular DNAs
catenated to each other. Each network consists of 40-50 maxicircles and 5000-10,000
minicircles. Te maxicircle DNA can be as long as 20-40kb. The minicircle DNA is about
645bp to 2500bp long. In some like L. tarantole maxicircle DNA include guide RNA
(gRNA) genes whose transcripts (thousands) associated with proteins (editosomes)
perform edi ng mRNAs either dele ng or addi on of Uridine.
Maxicircle DNA encodes for proteins needed for mitochondrial func on. Minicircle DNA
encode for guide RNAs which encode decode the encrypted maxicircle informa on.
Reproduc on of this kinetoplas d DNA takes place by disconnec ng these rings from
the parental kinetoplast DNA and subsequently join with the daughter Kinetoplast DNA.

Mitochondria very often found associated with smooth ER andCa2+ is transported into mitochondria.
www.mdpi.com;

Hypothe cal models of the role of contacts between mitochondria and ER in apoptosis; The hFis1/Bap31
pla orm transmits the mitochondrial stress signal to the ER via the ac va on of procaspase-8. The
cytosolic region of the ER integral membrane protein Bap31 is cleaved by ac vated caspase-8 to generate
proapopto c p20Bap31, which causes rapid transmission of ER calcium signals to the mitochondria via the
IP3 receptor. At close ER-mitochondria contact sites, mitochondria takes up calcium into the matrix via the
mitochondrial calcium channels MICU1 or LETM1. The massive influx of calcium leads to mitochondrial
fission, cristae remodeling, and cytochrome release. Mfn2 is enriched in the mitochondria-associated
membranes (MAM) of the endoplasmic re culum (ER), where it interacts with Mfn1 and Mfn2 on the
mitochondria to form inter-organellar bridges. Upon apoptosis signal, a BH3-only member of the Bcl-2
family, Bik, induces Ca2+ release from the ER and, in turn, induces Drp1 recruitment to the mitochondria
and their fragmenta on and cristae remodeling. SERCA- sarco /endoplasmic re culum Ca2+-ATPase.
MICU1, mitochondrial calcium uptake1. LETM1,leucine zipper/EF hand-containing transmembrane 1.
h p://www.hindawi.com/

Steps of mitochondrial fission and fusion; The figure highlights our current knowledge of the
mechanisms of fission and fusion in yeast (blue) and mammalian cells (red). For discussion, see text;h
p://physiologyonline.physiology.org/
Fission: The current model of fission in yeast is based on a trimer complex of Dnm1p, Mdv1p,
and Fis1p . The current model supports that Dnm1p translocates on ac va on to mitochondria,
where it func ons as a mechanoenzyme constric ng mitochondrial membranes like its homolog
dynamin I constricts the nascent endocyto c vescicle. In one model, a stable Fis1p-Mdv1p
complex is essen al to recruit Dnm1p, which then homo-oligomerizes. In a second model,
Dnm1p localiza on to mitochondria is independent of Fis1p and Mdv1p, which are conversely
responsible for the Dnm1p oligomeriza on step (FIGURE 2⇓). The protein Caf4p seems to have a
similar adaptor func on as Mdv1p. How mitochondrial fission is regulated remains obscure.
Remarkably, blockage of fission is not lethal to yeast, sugges ng that mitochondria can s ll
divide during cytokinesis as well as during meiosis and sporula on of diploid yeast cells.
Whether this is due to different protein machineries or to mechanical forces remains to be
elucidated.

Fusion: Fusion of mitochondria can be divided in at least three steps: docking, fusion of the
outer membrane, and fusion of the inner membrane. The trimeric complex of Fzo1p, Ugo1p,
and Mgm1p stands in the center of mitochondrial fusion. During docking, two or more Fzo1p on
juxtaposed mitochondria interact via their coiled-coil domains. A recently developed in vitro
assay showed that fusion of the outer membrane can be separated from that of the inner
membrane. Outer membrane fusion requires a pH gradient across the inner membrane and GTP,
whereas inner membrane fusion depends on membrane poten al and high levels of GTP (79)
(FIGURE 2⇑). Whether Mgm1p is responsible for the higher GTP consump on during inner
membrane fusion remains unclear. h p://physiologyonline.physiology.org/

h p://stmary.ws/

www.slideshare.ne

www.academic.brooklyn.cuny.edu

www.academic.brooklyn.cuny.edu

www.myscienceacademy.org

www.britannica.com; study.com

Mitochondria is involved in various func ons which are important to cell; www.edoc.hu-berlin.de

Central role of Kreb’s cycle that takes place in the mitochondrial matrix; www.ridge.icu.ac.jp;
www.gopixpic.com

Mitochondria are also involved in urea cycle and below citric acid cycle; edoc.hu-berlin.de

www.nutri onaloncology.org; www.phschool.com

Oxida ve phosphoryla on- Electron Transport chain

www.uic.edu

www.bmb.leeds.ac.uk

Detailed view of oxidative Phosphorylation; h p://employees.csbsju.edu/ biowiki.ucdavis.edu

Electron transport chain leads to produc on of ATP; h p://www.exatest.com/

www.ruf.rice.edu

www.biologicalphysics.iop.org

Mitochondrial Genome:
The size of mitochondrial genome varies from one species to the other, human
mitochondria contain ~16 Kbp and codes for13 proteins, 22tRNAs and 7 URFs. The
number of mit-DNAs per mitochondria can be 100 to 200 and the number of
mitochondria per cell varies from one to ten thousand or more. Refer to DNA replica on

www.genographic.nationalgeographic.com

Mitochondrial DNA rRNA genes and 13 protein coding genes and ~22 tRNA genes and 6-7 unknown URFs;
www.cellbiology.med.unsw.edu.au

Mitochondrial DNA Replica on sketch- D-lop mechanism;;

www.eplantscience.com; www.nar.oxfordjournals.org

Replica on of organelle DNA (Mitochondrial) with Replica on Ori-H andOri-L and Trnascrip onal start
sites-Hsp and Lsp-refer to DNA repica on

The mitochondrial genome. Jan Smei nk, Lambert van den Heuvel & Salvatore DiMauro; the outer and
inner circles represent Havy H and Lighter L strands. The D-loop contain Ori H and Ol on the inner strand.
Transcrip onal ini a on sites are shown as IT- H1 and IT-H2 and IT-L and the direc on of transcrip on
isindicated. mTERis mit transcrip on termina on factor binding sites. The 22 mRNA indicated by dots;
www.nature.com

Some gene c codes differ from universal coded, this is specific to mitochondrial codons

www.eplantscience.com

Mitochondrial in the process of division

Mitochondrial Protein Import:

Mitochondria require at least 1000 (yeast)-1500 (human data not consistent among
different published ar cles); proteins for its func on, but its genome codes for just 13
proteins in humans an d eight in yeasts and the rest of have to be imported from
cytosol. There is intricate communica on between mitochondria and nuclear genes
with respect to what proteins required by mitochondria and the same are transcribed by
the nuclear genome in response to mitochondrial signals; once they are synthesized
they are imported. The protein import can func on via signals between nucleus and
mitochondrial genomes. The import process is two-step process; one transport across
the outer membrane and then inner membrane. Interes ngly mitochondria are
connected to Endoplasmic Re culum (ER) membranes and it is involved in cellular ion
homeostasis and plays an important role in Apoptosis. Though mitochondria inherited
from bacteria, majority of the bacterial genes were transferred to host cell nucleus
(when?) and they func on each contribu ng to each other. Some of the transporters
involed are, TOM proteins (eight), Tim proteins (twelve), SAM three), Pam (two) several
others like Mdm, Mda, Hsp, Zim , heat shock proteins and some transporter are export
from mitochondria eg; Mss and Pnt1 (export from mitochondria) and few others.
Proteins that are imported have N-terminal sequences for the movement of proteins
from cytosol into mitochondrial peripheral space that is found between outer and inner
mitochondrial membranes. Some of the proteins imported into mitochondrial space are
further processed and get inserted to the inner surface of the inner membrane. Many
proteins are sorted in inner cristae membranes and some are found in free
mitochondrial space.

Principles of Mitochondrial Protein Biogenesis; http://www.sciencedirect.com/

(A) Cytosolic and mitochondrial protein synthesis. Most mitochondrial proteins are synthesized
on cytosolic ribosomes and are imported into the organelle. About 1% of mitochondrial proteins
are synthesized inside the organelle.
(B) Sorting pathways of mitochondria. The translocase of the outer membrane (TOM complex) is
the main entry gate into mitochondria. Subsequently, the precursor proteins follow different
sorting pathways. MIA, mitochondrial intermembrane space assembly; OXA, insertase/export
machinery of the inner membrane; SAM, sorting and assembly machinery; TIM22 complex,
carrier translocase of the inner membrane; TIM23 complex, presequence translocase of the inner
membrane. (Inset) The TOM complex consists of seven different subunits. The receptors Tom20,
Tom22, and Tom70 recognize precursor proteins and transfer them to the central component, the
channel-forming Tom40. Three small Tom proteins, Tom5, Tom6, and Tom7, are involved in the
assembly and dynamics of the TOM complex. (Presequence-carrying precursor proteins, red;
hydrophobic precursors with internal targeting signals, blue.); http://www.sciencedirect.com/

The Presequence Pathway to the Mitochondrial Inner Membrane and Matrix

(A) Forms of the presequence translocase of the inner membrane (TIM23 complex). (Left image)
Motor-free TIM23 complex that can insert preproteins into the inner membrane. (Right image)
TIM23 complex associated with PAM (presequence translocase-associated motor). The central
chaperone that binds to preproteins is mtHsp70. The function of mtHsp70 is regulated by the
cochaperones Pam16-Pam18, Tim44, Pam17, and the nucleotide exchange factor Mge1. The
function of the individual components is described in Table 1.
(B) Presequence-carrying preproteins are directed into the matrix by a sequential chain of
binding sites.
(C) Hypothetical model for the dynamic cooperation of the TIM23 complex of the inner membrane
(IM) with the TOM complex of the outer membrane (OM), the respiratory chain complexes III
(bc1-complex) and IV (cytochrome c oxidase, COX), and the motor PAM. The membrane
potential (Δψ) activates the Tim23 channel and exerts an electrophoretic effect on the positively
charged presequences of preproteins. ATP drives the action of mtHsp70. The mitochondrial
processing peptidase (MPP) removes the presequences.; http://www.sciencedirect.com/

Intermembrane Space Chaperones: The Carrier Pathway, and Machinery for Import and Assembly;
(A) Carrier pathway to the inner mitochondrial membrane. The noncleavable precursors of hydrophobic
metabolite carriers of the inner membrane are imported in several stages. Cytosolic chaperones guide the
precursor (stage I) to the receptor Tom70 (stage II). The precursor is transported through the translocase
of the outer membrane (TOM complex) in a loop forma on and interacts with the Tim9-Tim10 chaperone
complex (stage IIIa). Tim9-Tim10 guides the precursor through the intermembrane space (IMS) to the
carrier translocase of the inner membrane (TIM22 complex with bound Tim9-10-12) (stage IIIb). The
membrane poten al (Δψ) promotes inser on of the precursor into the inner membrane via the TIM22
complex (stage IV), followed by assembly into the mature form of the carrier protein (stage V).
(B) The machinery for import and assembly (MIA) of preproteins in the mitochondrial intermembrane
space is required for small IMS proteins with cysteine mo fs. The IMS receptor Mia40 binds to the
precursor via a transient disulfide bond. The sul ydryl oxidase Erv1 cooperates with Mia40 in the
oxida on of the precursor protein. Erv1 reoxidizes Mia40 and transfers electrons to cytochrome c (Cyt. c).
A third component, Hot13, assists in the oxida on of Mia40.
(C) Comparison of the disulfide relay systems of the mitochondrial intermembrane space, the
endoplasmic re culum, and the bacterial periplasm. In each system, disulfide bonds are introduced into
substrate proteins and electrons are removed (oxida on of substrate proteins). A disulfide genera ng
enzyme oxidizes a disulfide carrier that in turn oxidizes the substrate.
h p://www.sciencedirect.com/

Mitochondrial precursor proteins are synthesized in the cytosol and subsequently

imported into mitochondria. The import of mitochondrial intermembrane space
proteins is coupled with their oxida ve folding and governed by the mitochondrial
intermembrane space import and assembly (MIA) pathway. The cytosolic steps that
precede mitochondrial import are not well understood. We iden fied a role for the
ubiqui n-proteasome system in the biogenesis of intermembrane space proteins.
Interes ngly, the func on of the ubiqui n-proteasome system is not restricted to
condi ons of mitochondrial protein import failure. The ubiqui n-proteasome system
persistently removes a frac on of intermembrane space proteins under physiological
condi ons, ac ng as a nega ve regulator in the biogenesis of this class of proteins. Thus,
the ubiqui n-proteasome system plays an important role in determining the levels of
proteins targeted to the intermembrane space of mitochondria. h p://mcb.asm.org/
Mitochondria contain several hundreds of proteins. The mitochondrial content is
regulated by the uptake and degrada on of proteins. Stabiliza on of protein structure
by disulfide bonds was proposed to drive protein accumula on in the intermembrane
space of mitochondria. However, it remained unknown if structural altera ons could
lead to protein escape through the physiological barrier formed by the outer
mitochondrial membrane. In this work, we present evidence for size-dependent
retrograde movement of mitochondrial proteins to the cytosol. We iden fy the
translocase of the outer mitochondrial membrane channel protein Tom40 as an exit
route. Our results indicate that the retro-transloca on serves as an important
surveillance mechanism that regulates the abundance of intermembrane space proteins
in response to changes in cellular physiology.
The content of mitochondrial proteome is maintained through two highly dynamic
processes, the influx of newly synthesized proteins from the cytosol and the protein
degrada on. Mitochondrial proteins are targeted to the intermembrane space by the
mitochondrial intermembrane space assembly pathway that couples their import and
oxida ve folding. The folding trap was proposed to be a driving mechanism for the
mitochondrial accumula on of these proteins. Whether the reverse movement of
unfolded proteins to the cytosol occurs across the intact outer membrane is unknown.
We found that reduced, conforma onally destabilized proteins are released from
mitochondria in a size-limited manner. We iden fied the general import pore protein
Tom40 as an escape gate. We propose that the mitochondrial proteome is not only
regulated by the import and degrada on of proteins but also by their retro-transloca on
to the external cytosolic loca on. Thus, protein release is a mechanism that contributes
to the mitochondrial proteome surveillance. h p://www.pnas.org/

Protein transport across outer and inner mitochondrial membranes; Protein

import machinery; Protein Import Pathways of Mitochondria Most mitochondrial
proteins are synthesized as precursors in the cytosol and are imported by the
translocase of the outer mitochondrial membrane (TOM complex). (A) Presequence-
carrying (cleavable) preproteins are transferred from TOM to the presequence
translocase of the inner membrane (TIM23 complex), which is driven by the membrane
poten al (Dc). The proteins either are inserted into the inner membrane (IM) or are
translocated into the matrix with the help of the presequence translocase-associated
motor (PAM). The presequence are typically cleaved off by the mitochondrial processing
pep dase (MPP). (B) The noncleavable precursors of hydrophobic metabolite carriers
are bound to molecular chaperones in the cytosol and transferred to the receptor
Tom70. A er transloca on through the TOM channel, the precursors bind to small TIM
chaperones in the intermembrane space and are membrane inserted by the Dc-
dependent carrier translocase of the inner membrane (TIM22 complex). (C) Cysteine-
rich proteins des ned for the intermembrane space (IMS) are translocated through the
TOM channel in a reduced conforma on and imported by the mitochondrial IMS import
and assembly (MIA) machinery. Mia40 func ons as precursor receptor and
oxidoreductase in the IMS, promo ng the inser on of disulfide bonds into the imported
proteins. The sul ydryl oxidase Erv1 reoxidizes Mia40 for further rounds of oxida ve
protein import and folding. (D) The precursors of outer membrane b-barrel proteins are
imported by the TOM complex and small TIM chaperones and are inserted into the
outer membrane by the sor ng and assembly machinery (SAM complex). (E) Outer
membrane (OM) proteins with a-helical transmembrane segments are inserted into the
membrane by import pathways that have only been par ally characterized. Shown is an
import pathway via the mitochondrial import (MIM) complex. www.cell.com

https://www.biochemie.uni-freiburg.de
Most of the mitochondrial proteins are synthesized as precursor with cleavable N-end
cleavable sequences. The cleavable sequences for import are charged amphipatheic
alpha helices and they are recognized mitochondrial membrane bound receptors on the
outer membrane (TOM). A er transloca on into perimitochondrial space the
preproteins transferred to inner membrane translocator protein complexes (TIM23). The
presequence associated motor protein (PAM) drives the preproteins are translocated
into mitochondrial matrix. Once translocated the presequences are cleaved in the
matrix. Some cleavable proteins get arrested by TIM23 and later they are released into
inner membrane. In some proteins are translocated into matrix and followed by
inser on into inner membrane matrix

www.cc.scu.edu.cn; cc.scu.edu.cn

www.cc.scu.edu.cn; cc.scu.edu.cn

www.biochemie.uni-freiburg.de

Movement of Mitochondrai:
Mitochondrial movement in a cell, where some cells are long reaching several
micrometers from the cell body ex. Axons of nerve cells. Most of the movements are
anterograde example from the cell base towards the synaptic surface of the neuronal cell
and the backward movement is called retrograde. Anterograde movement is from the
base or cell body of the cells to toward microtubule plus end and they use Kinesin motor
proteins. Retrograde transport is from plus end of MT to negative end or synaptic surface
to basal cell body. Similarly many other cellular components are transported forward and
backward.

http://jcs.biologists.org/

http://bio100.class.uic.edu/

Important structures involved are microtubules which use motor proteins Kinesins
forward (anterograde) to microtubule (MT) plus ends and Dyneins are used to transport
from plus ends towards minus ends called retrograde movement. Mitochondria are also
transported in axon by the same process. New mitochondria generated in cell bodies are
transported to the nerve ends.

http://jcs.biologists.org/

Axonal transport from neuron cell body to terminal region by retrograde movement of
endosomes all along the length of microtubule from minus end toward plus ends. Actins
are also involved in mitochondrial transport using myosin. https://publish.illinois.edu;
http://file script.org

Mitochondrial changes in neuro-degenera on; Most neurodegenera ve diseases result in defects in

oxida ve phosphoryla on, respiratory dysfunc on, increased ROS produc on, lowered mitochondrial
membrane poten al, decrease in synthesis of an oxidants, mtDNA muta ons, impaired protein import,
increased fragmenta on of mitochondria, and ac va on of mitophagy and apoptosis. Sources of reac ve
oxygen species are marked by red stars. Abbrevia ons: H , Hun ng n; Ub, ubiqui n; ΔΨ, membrane
poten al; MPTP, 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine; ROS, reac ve oxygen species; mtDNA,
mitochondrial DNA; Aβ, amyloid beta; APP, amyloid precursor protein; TIM, translocase of inner
membrane; TOM, translocase of outer membrane; SOD1, superoxide dismutase 1; VDAC, voltage
dependent anion channel; ANT, adenine nucleo de translocator; Pink1, PTEN-induced puta ve kinase
1; αKGDH, alpha ketoglutarate dehydrogenase; cyt c, cytochrome c.;ww.hindawi.com.
For More Informa on Read-www.grkraj.org

Uniparentall Inheritance of mitochondria in various groups of plant and animal

systems:
Uniparental inheritance of mtDNA is observed in many sexually reproducing
species. However, it may be accomplished by various different strategies in
different species. Possible mechanisms are listed as below: 1)decrease in the
content of mtDNA during spermatogenesis,2) elimination of mtDNA from mature
spermatozoa, 3) prevention of sperm mitochondria from entering the oocyte, 4)
active degradation of the paternal mtDNA in the zygote, 5) selective degradation
of the whole paternal mitochondria in the zygote.

Size of mitochondrial DNA varies; http://www.sciencedirect.com/

Fate of Mitochondria found in Spermatazoids:

Mitochondria are inherited by mother and not by father; it is always maternal
inheritance. Any sperm that enters fer lized egg, the sperm mitochondria is degraded
due to a tag that is carried during spermatogenesis. However mitochondrial popula on
is same in all for it lacks proper repair mechanism and proof reading capabili es.
Muta on rate of mitochondrial DNA is ten mes higher than nuclear DNA. More than
30 ubiqui na ng enzymes have been iden fied as important regulators of
spermatogenesis.

Morphological changes during spermiogenesis. In elongated

spermatozoids mitochondria are aligned along the anterior part of the
flagellum and ghtly packed to form the helically arranged mitochondrial
sheath; h p://www.mdpi.com/

Mature sperm cell is slender; in the middle part , the mitochondria are thick and rig
shaped. The DNA of the nucleus is maximally condensed; Mitochondria present in mid
piece get destroyed by using ubiquitinated surfaces by proteasomes,
Neck contains two centrioles, Mid-piece consists of a sheath of ring-shaped
mitochondrial grouped, the principal piece has a sheath of a ring of fibers around
axoneme, The tail consists of only the 9+2 structure of the axoneme, The mature sperm
cell is approximately 60um long and completely enveloped by the plasma membrane.

1.Plasma membrane, 2. Outer acrosomal membrane, 3.Acrosome, 4. Iner acrosomal

membrane, 5.Nucleus, 6. Proximal centriole, 7. Rest of the distal centriole, 8. Thick
longitudinal fibers, 9. Mitochondria, 10. Axoneme, 11. Annulus, 12. Ring fibers; A.Head,
B. Neck, D. Principal Piece, and E.End piece. http://www.embryology.ch/

http://leavingbio.net/
Maternal inheritance of mitochondrial DNA has long been regarded as a major paradox in
developmental biology. While some confusion may still persist in popular science, research data
clearly document that the paternal sperm-borne mitochondria of most mammalian species enter
the ooplasm at fertilization and are specifically targeted for degradation by the resident ubiquitin
system. Ubiquitin is a proteolytic chaperone that forms covalently linked polyubiquitin chains on
the targeted proteinaceous substrates. The polyubiquitin tag redirects the substrate proteins to a
26-S proteasome, a multi-subunit proteolytic organelle. Thus, specific proteasomal inhibitors
reversibly block sperm mitochondrial degradation in ooplasm. Lysosomal degradation and the
activity of membrane-lipoperoxidating enzyme 15-lipoxygenase (15-LOX) may also contribute to
sperm mitochondrial degradation in the ooplasm, but probably is not crucial. Prohibitin, the major
protein of the inner mitochondrial membrane, appears to be ubiquitinated in the sperm
mitochondria. Occasional occurrence of paternal inheritance of mtDNA has been suggested in
mammals including humans. While most such evidence has been widely disputed, it warrants
further examination. Of particular concern is the documented heteroplasmy, i.e. mixed mtDNA
inheritance after ooplasmic transplantation. Intracytoplasmic sperm injection (ICSI) has inherent
potential for delaying the degradation of sperm mitochondria. However, paternal mtDNA
inheritance after ICSI has not been documented so far. http://www.ncbi.nlm.nih.gov/

h p://image.slidesharecdn.com/

http://image.slidesharecdn.com/fertilization notes

Mul ple mechanisms of paternal mtDNA elimina on.

In mammals, paternal mitochondria are modified by ubiqui n binding to specific
proteins on mitochondria during spermatogenesis and degraded by the
proteasomes and/or lysosomes a er fer liza on. (b) In Caenorhabdi s elegans,
paternal mitochondria and membranous organelles (MOs) are engulfed by
autophagosomes and targeted for lysosomal degrada on a er fer liza on.
Ubiqui na on is detectable on MOs. (c) In Oryzias la pes, the number of mtDNA
nucleoids decreases during spermatogenesis. A er fer liza on, paternal mtDNA
is further degraded before the destruc on of the mitochondrial structure. (d)
In Drosophila melanogaster, paternal mtDNA is degraded by EndoG during
spermatogenesis. In EndoG mutants, the remaining mtDNA nucleoids are
eliminated by investment cones and deposited into a waste bag. (e) In the
isogamous species of Physarum polycephalum and Chlamydomonas reinhard i,
mtDNA from 1 parent is selec vely degraded in the mitochondria a er gamete
fusion.

Uniparental inheritance of mtDNA, as well as chloroplast DNA (cpDNA) or plas d DNA (ptDNA), is
also observed in plants ... However, in most mammals, including humans and mice, the paternal
mitochondria and their mtDNA do enter the oocyte cytoplasm upon fer liza on; In mammals
paternal mitochondria are ubiqui n mediated degrada on takes place in the fer lized egg.

cpDNA is maternally inherited in the four-o'clock plant (Mirabilis jalapa) . In Sequoia

sempervirens, in which mtDNA and cpDNA are paternally inherited. n the geranium
plant (Pelargonium zonale), cpDNA is inherited maternally, paternally or biparentally .
When zygotes receive cpDNA from both parents, cpDNAs from each parent are rapidly
segregated following cell division, resul ng in the mature plant inheri ng clonal sectors
of cells that are homoplas c for only 1 type of cpDNA. In angiosperm species, such
as Arabidopsis thaliana, both mtDNA and ptDNA are maternally inherited and . In A.
thaliana, the ac ve degrada on of paternal mtDNA and ptDNA takes place during pollen
development.

The content of mtDNA and cpDNA gradually decreases during meiosis, and these DNAs are no
longer detectable in mature pollen. The wild-type DPD1 encodes a pollen-specific Mg2 +-
dependent exonuclease, which is transported into both chloroplasts and mitochondria,
sugges ng a direct role for this exonuclease in the degrada on of mtDNA and ptDNA.
h p://www.sciencedirect.com/; h p://biology-themiracleoflife.blogspot.in/; h p://www.ncbi.nlm.nih.gov/