Charmaine and Gabi
Charmaine and Gabi
Charmaine and Gabi
I-2 Sugar
II.1 Dinucleotides
II.3.4 G4-DNA
IV Various
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Deoxyribonucleic acid (DNA) IS the genetic information of most living organisms (a
contrario, some viruses, called retroviruses, use ribonucleic acid as genetic
information).
- DNA can be copied over generations of cells: DNA replication
- DNA can be translated into proteins: DNA transcription into RNA, further translated
into proteins ,
- DNA can be repaired when needed: DNA repair .
Ribonucleic acids (RNAs) are described in another chapter ( mRNA, r-RNA, t-
RNA... )
I-2 Sugar:
Deoxyribose, which is a cyclic pentose (5-carbon sugar). Note: the sugar in RNA is a
ribose. Carbons in the sugar are noted from 1' to 5'. A nitrogen atom from the
nitrogenous base links to C1' (glycosidic link), and the phosphate links to C5' (ester
link) to make the nucleotide. The nucleotide is therefore: phosphate - C5' sugar C1' -
base.
Note: other nitrogenous bases exist, in particular methylated bases derived from the
above mentioned; methylation of the bases has a functional role (see chapter ad hoc).
Glossary:
- Nucleoside names: deoxyribonucleosides in DNA: deoxyadenosine,
deoxyguanosine, deoxycytidine, deoxythymidine in DNA (ribonucleosides in RNA:
adenosine, guanosine, cytidine, uridine).
- Nucleotide names: deoxyribonucleotides in DNA: deoxyadenylic acid,
deoxyguanylic acid, deoxycytidylic acid, deoxythymidylic acid (ribonucleotides in
RNA: adenylic acid, guanylic acid, cytidylic acid, uridylic acid).
II.1 Dinucleotides
DNA is made of two ("duplex DNA") dextrogyre (like a screw; right-handed) helical
chains or strands ("the double helix"), coiled around an axis to form a double helix of
20A° of diameter. The two strands are antiparallel (id est: their 5'->3' orientations are
in opposite direction). The general appearance of the polymere shows a periodicity of
3.4 A°, corresponding to the distance between 2 bases, and another one of 34 A°,
corresponding to one helix turn (and also to 10 bases pairs).
II.2.1 Hydrogen bounds: bases pairing
The (hydrophobic) bases are stacked on the inside, there planes are perpendicular to
the axis of the double helix. The outside (phosphate and sugar) are hydrophilic.
Hydrogen bounds between the bases of one strand and that of the other strand hold the
two strands together (dashed lines in the drawing).
A purine on one strand shall link to a pyrimidine on the other strand. As a corollary,
the number of purines residues equals the number of pyrimidine residues.
A binds T (with 2 hydrogen bounds).
G binds C (with 3 hydrogen bounds: more stable link: 5.5 kcal vs 3.5 kcal).
Note: the content in A in the DNA is therefore equal to the content in T, and the
content in G equals the content in C.
This strict correspondance (A<->T and G<->C) makes the 2 strands complementary.
One is the template of the other one, and reciprocally: this property will allow exact
replication (semi-conservative replication: one strand -the template- is conserved,
another is newly synthesized, same with the second strand, conserved, allowing
another one to be newly synthesized; see chapter ad hoc).
Notes:
Hydrogen bounds in base pairing are sometimes different from the model of Watson
and Crick above described, using the N7 atom of the purine instead of the N1
(Hoogsteen model).
II.2.2 Major groove and minor groove
The double helix is a quite rigid and viscous molecule of an immense length and a
small diameter. It presents a major groove and a minor groove.
The major groove is deep and wide, the minor groove is narrow and shallow.
DNA-protein interactions are major/essential processes in the cell life (transcription
activation or repression, DNA replication and repair).
Proteins bind at the floor of the DNA grooves, using specific binding: hydrogen
bounds, and non specific binding: van der Waals interactions, generalized electrostatic
interactions; proteins recognize H-bond donnors, H-bond acceptors, metyl groups
(hydrophobic), the later being exclusively in the major groove; there are 4 possible
patterns of recognition with the major groove, and only 2 with the minor groove (see
iconography).
- Some proteins bind DNA in its major groove, some other in the minor groove, and
some need to bind to both.
Notes:
- The 2 strands are called "plus" and "minus" strands, or "direct" and "reverse"
strands. At a given location where one strand (any of the two) bears coding sequences,
it is unlikely (but not impossible) that the other strand also bears coding sequences.
- DNA is ionized in vivo and behave like a polyanion.
The double helix as described above is the "B" form of the DNA; it is the form the
most commonly found in vivo, but other forms exist in vivo (see below) or in vitro.
The "A" form resemble B-DNA but it is less hydrated than B-DNA, "A" form is not
found in vivo.
DNA is a molecule which moves, fidgets, does gymnastics, dances. The structures
below cited are being proved to have funtional roles; on the other hand, they may
favour DNA breaks and further deletions, amplification, recombination, and
mutations.
Glossary:
Palindromes: these are names that read the same backwards and forwards (e.g. "DNA
LAND"). DNA uses to play with palindromes: see below).
II.3.1 Z-DNA
- Z form is a levogyre (left handed) double helix with a zig-zag conformation of the
backbone (less smooth than B-DNA). Only one groove is observed, resembling the
minor groove, the base pairs being set off to the side, far from the axis. The bases
(which form the major groove -close to the axis- in B-DNA) are here at the outer
surface. Phosphates are closer together than in B-DNA. Z-DNA cannot form
nucleosomes.
- A high G-C content favours Z conformation. Cytosine methylation, and molecules
which can be present in vivo such as spermine and spermidine can stabilize Z
conformation.
- DNA sequences can flip from a B form to a Z form and vice versa: Z-DNA is a
transient form in vivo.
- Z-DNA formation occurs during transcription of genes, at transription start sites near
promoters of actively transcribed genes. During transcription, the movement of RNA
polymerase induces negative supercoiling upstream and positive supercoiling
downstream the site of transcription The negative supercoiling upstream favours Z-
DNA formation; a Z-DNA function would be to absorb negative supercoiling. At the
end of transcription, topoisomerase relaxes DNA back to B conformation.
- Certain proteins bind to Z-DNA, in particular double-stranded RNA adenosine
deaminase (ADAR1), a Z-DNA binding nuclear-RNA-editing enzyme; this enzyme
converts adenine to inosine in the pre-mRNA. Following, ribosomes will interpret
inosine as guanine, and the protein coded with this epigenetic modification will be
different (see chpater on Epigenetics).
Notes:
- Z-DNA antibodies are found in lupus erythematosus and other autoimmune diseases.
- Double stranded RNA (dsRNA) can adopt a Z conformation.
DNA is associated with proteins: histones and non histone proteins, to form the
chromatin. DNA as a whole is acidic (negatively charged) and binds to basic
(positively charged) proteins called histones: see chapter Chromatin
There is 3 x 10 9 nucleotide pairs in the human haploid genome representing about 30
000 genes dispersed over 23 chromosomes for a haploid set.
IV Various
- DNA is found in the nucleus of the cell, but a small amount is also present in the
mitochondria.
- Mitochondrias would originate from archeobacterias which became endosymbiotic
to eukaryotic cells.
- Their genetic code is different from the so-called "universal" code (UGA, AUA,
AGA, AGG: respectively STOP, Ile, Arg, Arg in the universal code, and Trp, Met,
STOP, STOP in the mitochondria of mammals, and other meanings in mitochondria
of other spieces).
- The number of DNA copies in one given mitochondria is variable.
- Mitochondrial DNA is circular, with a heavy and a light chains, has no introns, not
any non-coding sequence.
- Genes from the mitochondria code for proteins involved in electron transport,
ribosomic RNAs (rRNAs), and transfer RNAs (tRNAs).
- Each DNA strand is transcribed, then cut into the mRNAs, but also into rRNAs and
tRNAs.
Note: the mitochondria also use proteins imported from the cytoplasm of the cell (and
coded by the nucleus); so far, proteins from the mitochondria are not exported into the
cytoplasm except in case ofapoptosis.
The double helix undergoes unwinding in vitro with heat, extremes ph, and other
conditions (urea, ...). A melting point can be calculated; it is characteristic of the A/T
versus G/C proportion of the specimen studied, due to the fact that there is only 2
hydrogen bounds in A/T, and 3 in G/C, a more stable binding. Upon denaturation, the
physical properties of the DNA change; e.g. hyperchromic effect: light absorption at
260 nm is higher with denatured DNA than with double standed DNA. Light
absorption also varies according to the A/T vs G/C proportion: it is higher in A/T rich
specimens than in G/C rich ones.
DNA denaturation is to be known, because: 1- it allows to measure A/T vs G/C
content; 2- it is the basis of in situ hybridization techniques (see Methods in Genetics).