Regulation of Gene Expression

in Eukaryotes

Presented by
Quratulain (19-Arid-1403)
Asadullah(19-Arid-1379)
 Levels of Gene Expression
 Genome level
 Transcription
 Post transcriptional
modification
 Translation
 Genome Level
 Chromatin structure
DNA in eukaryotic cells is extensively folded into protein-DNA complex
called chromatin.
Chromatin Heterochromatin.
A lesser coiled trans critically A highly condensed transcriptically
Active regions which can be inactive region. The genes in this
Easily accessed by RNA region can not be accessed by RNA
Polymerases polymerases for active transcription
 Histone modifications-these modifications make a
region of gene either transcriptically active or inactive.
(a) Acetylation
Acetylation--- condensation of DNA----
transcription of genes in that region
 Histone methylation
 Methylation of arginine residue at the 4th position of histone
H4-
 Opens the chromatin structure
 Leading to transcriptional activity
 Methylation of lysine residues at the 4th and 79th position of
histone H3
 Opens the chromatin structure
 Leading to transcriptional activation
 DNA-methylation

 Methylation of lysine residues at the 9th and 27th

position of histone H3
 Condenses the chromatin structure
 Leading to transcriptional inactivation
2. Transcription level

 INTRODUCTION
 Transcription has been defined in various
ways. Some definitions of transcription are given here.
 The synthesis of RNA from a single strand of a DNA
molecule in the presence of enzyme RNA polvmerase is
called transcription.
 In other words, the process of formation of a messenger
RNA molecule using a DNA molecule as a template is
referred to as transcription.
1)SYNTHESIS
 RNA is synthesized from a DNA template.
 The RNA is processed in to messenger RNA ( m
RNA) which is then used for synthesis of a protein.
 The RNA thus synthesized is called m RNA , because it
carries a genetic message from the DNA to the protein
synthesizing machinery of the cell.
 The main difference between RNA and DNA sequence is
the presence of U, or uracil in RNA instead of the T of
thymine of DNA.
2)TEMPLATE USED

 • The RNA is synthesized from a single strand or

template of a DNA molecule,
 • The stretch of DNA that is transcribed into an
RNA molecule is called a transcription unit.
 A transaction unit codes the sequences that is
translated into protein.
 • It also directs and regulates protein synthesis.
 The DNA strand which is used in RNA synthesis
is called template strand ; because it provides
the template for ordering the sequence of
nucleotides in an RNA transcript.
 
 The DNA strand which does not take part in DNA
synthesis is called coding strand because
it's nucleotide sequence is the same as that of the
newly created RNA transcript.
3)ENZYME INVOLVED
 The process of transcription is catalyzed by the
specific enzyme called RNA polyrnerase. 
 • DNA sequence is enzymatically copied by RNA
polymerase to produce a complement tarry
nucleotide RNA strand.
 • In eukaryotes , there are three classes of RNA
polymera5e I, II and III which are involved in the
transcription of all protein genes.
 4)GENETIC
INFORMATION COPIED
 In this process the genetic information coded in DNA is
copied into a molecule of RNA. 
 The genetic information is transcribed or copied from DNA
to RNA. 
 In other words it results in the transfer of genetic
information from DNA into RNA
 5)FIRST STEP

 The expression of a gene consists of two major

steps vise ; transcription and translation. 
 Thus transaction is the first step in the process of
gene regulation or protein synthesis.
6)DIRECTION SYNTHESIS
 As in DNA replication , RNA is synthesized in the 5' -3'
direction.
 The DNA template strand is read 3'-5' by RNA polymerise and
the new RNA strand is synthesized in the 5' -3' direction.
 RNA polymerase bind to the 3' end of a gene on the DNA
template strand and travels to ward the 5' end.
 The regulatory sequence that is before or 51 of the coding
sequence is called 5' untranslated region and sequence found
following or 31 of the coding sequence is called 3’
untranslated region.
 MECHANISM OF
TRANSCRIPTION
 The mechanism of transcription consists of three
major steps :
 1. Initiation
 2. Elongation 
 3. Termination
 1)INITITATION
 Pre - initiation
 In initiation of transcription does not require a
prime to start.
 RNA polymerase simply binds to the DNA and
along with other cofactors . the DNA to create an
initiation bubble so that the RNA polymerase has
access to the single stranded DNA template.
 However, RNA polymerase does require a
promoter like sequence.
 PROXIMAL CORE
PROMOTERS
 TATA promoters are found around —30bp to the
start site of transcription. 
 Not all genes have TATA box promoters and there
exists TATA -less promoters and there exists
promoter consensus sequence is TATA.
 INITIATION
 • In eukaryotes and archaea, transcription initiation is
Far more complex.
 • The main difference is that eukaryotic polyrnerases do
not recognize directly their core promoter sequence.
 • In eukaryotes, a collection of proteins called
transcription factors mediate the binding of RNA
polymerase and the initiation of transcription.
 • Only after attachment of certain transcription factors
to the promoter , the RNA polymerase binds to it.
 • The complete assembly of transcription factors
and RNA polymerase bind to the promoter,
called transcription initiation complex.
 • Initiation starts as soon as the complex is
opened and the first phosphodiester bond is
formed.
 This is the end of Initiation.
 • RNA poi II does not contain a subunit similar to
the prokaryotic factor, which can recognize the
promoter and unwind the DNA double helix.
 In eukaryotes ,these two functions are carried out by a
set of proteins called general transcription factor.
 The RNA pol II is associated with six general
transcription factors, designated as TFIIA, TFIIB,
TFIIF and TFIIH where TF stands for for transcription
factors and II for the RNA pol II.
 TTFIID consists of TBP (TATA box binding protein)
and TAFS (TBP associated factors).
 The role of TBP is the core promotor.
 • TAFs may assist TBP in this process.
 • In human cells TAFs are formed by 12 subunits.
 IF One of them TAFs 250 ( with molecular
weight 2501(0
has the histone acetyl transfers activity which
can relieve the binding between DNA and histone
in the nucleosome.
 • The transcription factor which catalyzes DNA
melting is TF1111.
 • However, before TFIIH can unwind DNA the
RNA pol III at feast five general transcription
factors have to form a pre - initation complex.
PROMOTER CLEARENCE
 After the bond is synthesized that RNA
polymerase must clear the promotor.
 during this time there is a tendency to release
the RNA transcript and produce truncated
transcripts.
 This is called abortive imitation and is common
for botj eukaryotes and prokaryotes.
 Once the transcripts reaches approximately 23
nucleotides it is no longer slips and elongation
can occur.
 This is an ATP dependent process.
 2)ELONGATION
 • For RNA synthesis one strand of DNA known as
the template strand or non coding strand is used as
a template.
 As transcription proceeds RNA polymerase
traverses the template strand and uses base
pairing complementarity with the DNA template
to create an RNA copy.
 • Although RNA polymerase traverses the
template strand from 31-5' the coding strand is
usually used as the reference point 50
transcription is said to go from 51-3P.
 This produces an RNA molecules from 5’-3’ and
exact copy of the coding strand except that
thyamin are replaced uracils and the nucleotides
are composed of a ribose sugar where DNA has
deoxyribose in its sugar phosphate backbone.
 After pre- initiation complex is assembled at the
promoter TFIIH can use its helicase activity to
unwinds DNA.
 This requires energy released from ATP
hydrolyses.
 The DNA melting start from about 10bp.
 Conti….
 Then RNA pol II uses nucleoside triphosphate to
synthesize a RNA transcripts.
 During RNA elongation TFIIF, remains attached to
the RNA polymerase but all of the other
transcription factors have dissociative from P IC .
 The carboxyl terminal domain(CTD) of the largest
subunit of RNA pol II is critical for elongation
 In the imitation phase CTD is un phosphorylated
but during elongation it has to be phosphorylated.
 The domain contains many proline, serine residues.
 3)TERMINATION

 In eukaryotic transcription the mechanism of termination is

not very clear.
 • In other words, it is not well understood.
 • It involves cleavage of the new transcript followed by
template independent addition of as at its new 3' and in a
process called polyadenylation.
 • Eukaryotic protein genes contain a poly -A signal located
down stream of the last exon.
 • This signal is used to add a series of adenylate residues RNA
processing.
 • Transcription often terminates at 0.5 - 2 kb downstream of
the poly A signal.
TRANSLATION LEVEL

 Initiation: The first phase of translation

 1 Translation begins when mRNA attaches to
30S.
 tRNA comes and binds to mRNA where
nucleotide code matches.
 1 This triggers 50S binding to 30S. 50S is where
all
tRNA's will bind.
 Elongation
 The second phase
 Two binding sites on 50S. A site and P site ,which aid
in continuing translation.
 First tRNA connected at A site .Now moves to P site
as another tRNA approaches.
 Second tRNA binds to A site.
End Of Translation
 Ribosomes was moving along nucleotide triplets one by
one
 Ribosomes reaches ‘Stop codon’ peptide chain finished.
Last tRNA. leaves ribosomes leaving behind complete
peptide chain.
 Ribosomes separates from mRNA
 Ribosome subunits also separate, and will remain this
way until another mRNA comes along to restart the
process.