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Transcription_
-Eukaryotes
GP-501 ; Dated: 22-10-2013
Asif B Shikari
ProteinRNADNA
Genetic information from DNA is transcribed to RNA and that
information from RNA is translated to polypeptide chain. This
concept of flow of biological information from DNA to RNA and
then to protein was given by Crick and is known as “Central
Dogma”. This central Dogma holds true for those organisms in
which DNA is genetic material.
TranslationTranscription
Central dogma
Self replication
Classes of genes transcribed by
eukarytoic RNA polymerase
Basic elements in Transcription
Classes of Transcriptional Factors
Outline of
Eukarytoic transcription
First Step in eukarytoic transcription
Pre-initiation complex
 TBP uses
extensive β-
sheet to
recognize the
minor groove of
TATA element.
 A:T base pairs
are favoured
because they
are more readily
distorted to
allow the initial
opening of the
minor groove.
 Also interactions
between P in
backbone and
basic residues in
β-sheet stabilize
association.
 TBP-TATA
binding means
specificity
Out of 10 TAFs, two bind at Inr element and
Downstream Promoter element
 TFIIB interacts with TBP and also with DNA at BRE
 Start site selection: [TFIIB mutants result in shift of transcription start
site]
 Results in Uni-directional transcription
 Pol II + TFIID +TFIIB form minimal initiation complex
 Functions in TFIIF recruitment
Tight Binding
TFIID RNAP
Elongation Phase Starts
Phosphorylation of the
polymerase could be
the trigger that allows
the polymerase to shift
from initiation
to elongation mode.
 Kinase P-TEFb is recruited to Polymerase by Transcriptional
activators
 Once bound, this enzyme phosphorylates Ser at Position 2 of
CTD repeats.
 P-TEFb phosphorylates hSTP5 protein and activates it. hSTP5
after activation leads to elongation.
 P-TEFb phosphorylates TAT-SF1 elongation factor.
 TFIIS is another factor which is recruited and stimulates
elongation and proof reading.
 Simultaneously, at the time of elongation, the enzymes for 5’
capping and polyadenylation are recruited at the
corresponding sites.
Elongation Phase continues
Prokaryotes Eukaryotes
A single RNA polymerase;
RNA Pol
Multiple RNA Polymerases;
Pol I, Pol II, Pol III
RNA polymerase binds
directly to Promoter
sequences
First GTFs bind to Promoter
& then binds Pol II (Pre-
initiation complex)
Sigma as specificity factor GTFs as specificity factors
Polycistronic message Transcription of single gene
Translation and
transcription is coupled
Transcription (Nucleus) and
Translation (Cytosol) are
independent
No definite phase of its
occurence
Occurs in G1 and G2 phase
of cell cycle
Comparing both
Prokaryotes Eukaryotes
RNAs are released and
processed in cytoplasm
RNAs processed in nucleus
Initiation doesnt need any
initiation factors
Initiation requires TFs
RNA Pol is complex of 5
polypeptides
RNA Pol is complex of 10-15
polypeptides
Little or no processing of
RNA
Capping, Splicing &
Polyadenylation takes place
Thankyou
all

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Transcription eukaryotes

  • 1. Transcription_ -Eukaryotes GP-501 ; Dated: 22-10-2013 Asif B Shikari
  • 2.
  • 3. ProteinRNADNA Genetic information from DNA is transcribed to RNA and that information from RNA is translated to polypeptide chain. This concept of flow of biological information from DNA to RNA and then to protein was given by Crick and is known as “Central Dogma”. This central Dogma holds true for those organisms in which DNA is genetic material. TranslationTranscription Central dogma Self replication
  • 4.
  • 5. Classes of genes transcribed by eukarytoic RNA polymerase
  • 6. Basic elements in Transcription
  • 9. First Step in eukarytoic transcription Pre-initiation complex
  • 10.  TBP uses extensive β- sheet to recognize the minor groove of TATA element.  A:T base pairs are favoured because they are more readily distorted to allow the initial opening of the minor groove.  Also interactions between P in backbone and basic residues in β-sheet stabilize association.  TBP-TATA binding means specificity Out of 10 TAFs, two bind at Inr element and Downstream Promoter element
  • 11.
  • 12.  TFIIB interacts with TBP and also with DNA at BRE  Start site selection: [TFIIB mutants result in shift of transcription start site]  Results in Uni-directional transcription  Pol II + TFIID +TFIIB form minimal initiation complex  Functions in TFIIF recruitment
  • 13.
  • 14.
  • 15.
  • 16.
  • 18.
  • 20. Phosphorylation of the polymerase could be the trigger that allows the polymerase to shift from initiation to elongation mode.
  • 21.  Kinase P-TEFb is recruited to Polymerase by Transcriptional activators  Once bound, this enzyme phosphorylates Ser at Position 2 of CTD repeats.  P-TEFb phosphorylates hSTP5 protein and activates it. hSTP5 after activation leads to elongation.  P-TEFb phosphorylates TAT-SF1 elongation factor.  TFIIS is another factor which is recruited and stimulates elongation and proof reading.  Simultaneously, at the time of elongation, the enzymes for 5’ capping and polyadenylation are recruited at the corresponding sites. Elongation Phase continues
  • 22. Prokaryotes Eukaryotes A single RNA polymerase; RNA Pol Multiple RNA Polymerases; Pol I, Pol II, Pol III RNA polymerase binds directly to Promoter sequences First GTFs bind to Promoter & then binds Pol II (Pre- initiation complex) Sigma as specificity factor GTFs as specificity factors Polycistronic message Transcription of single gene Translation and transcription is coupled Transcription (Nucleus) and Translation (Cytosol) are independent No definite phase of its occurence Occurs in G1 and G2 phase of cell cycle Comparing both
  • 23. Prokaryotes Eukaryotes RNAs are released and processed in cytoplasm RNAs processed in nucleus Initiation doesnt need any initiation factors Initiation requires TFs RNA Pol is complex of 5 polypeptides RNA Pol is complex of 10-15 polypeptides Little or no processing of RNA Capping, Splicing & Polyadenylation takes place