EUKARYOTIC

TRANSCRIPTION
Features
 Multiple RNA polymerases transcribe distinct classes of genes

 RNA polymerase need to interact with a variety of proteins

 Takes place on chromatin

 Undergo post transcriptional modification

 The modified mRNA has a 5’ methylated cap and a 3’ poly A tail

 Eukaryotic mRNAs are monocistronic

 Introns are spliced during modifications

 Many mRNA are long lived

 Eukaryotic RNA polymerases
 3 distinct polymerases are there

RNA polymerase I – transcribe most rRNA genes ( 28s, 18s &

5.8s), resistant to α - amanitin

RNA polyemerase II – transcribe all protein coding genes and

some small nuclear RNA genes, present in nucleoplasm,
sensitive to α – amanitin

RNA polymerase III – transcribes genes for tRNA , 5s rRNA,

small RNAs involved in protein transport and splicing, present
in nucleoplasm and moderately sensitive to α - amanitin
 All the 3 are made up of 12-17 subunits
 They have 9 conserved sequences
Selection of site : the promoter
 3 specific regions between 25-100bp upstream the
initiator site

 Goldberg – Hogness box : TATA box, 20-30 bp

upstream to initiation site, sequence TATAAAT

 CAAT box : 70 – 80bp upstream

 GC box : 60 – 100bp upsream, GGGCGC is the

sequence,

 Initiator or Inr sequence and down stream

promoter element 30bp upstrem to transcription
initiation site
General transcriptional factors and
transcription initiation by RNA pol II

 About 10% of of human genome encodes

for transcription factors
 5 transcriptional factors are required for
transcription by RNA pol II
 General transcription factors + RNA pol II
-- pre – initiation complex or
transcription complex
 Steps : Initiation
 Binding of Transcription factor ( TF ) – TF II D to the promoter
region

 TF II D composed of multiple subunits called TATA binding

proteins ( TBP ) and TBP associated factors (TAF)

 TBP binds to TATA box whereas other subunits attach to Inr and
DPE

 TF II D is associated with inhibitory factors such as DR1 and DR2

 TF II A binds to TF II D to prevent binding of inhibitors and to

stabilize TF II D – DNA complex

 TF II D followed by TF II B in association with TBP, act as bridge

between DNA and RNA pol with the help of TF II F
 Addition of 2 factors TF II E & TF II H completes the formation
of initiation complex

 TF II H functions as helicases to unwind DNA around initiation site

and one of its subunit is protein kinase which will phosphorylate
repeated sequence at C – terminal domain (CTD) of pol II

 This will release the Pol from pre initiation complex and recruit
other polymerases to initiate transcription

 Other additional factors called large protein complex mediators

are also involved

 During the phosphorylation reaction TFs shed from RNA pol and
recovery of Abortive initiation occur

 Then elongation phase starts

Elongation
 Transcription elongation factors are involved

 2 factors

 TF II F – RNA chain growth

 TF II S – elongation of RNA chain, clears the obstructions by

hydrolyitic cleavage at 3’ end of RNA chain to help in forward
movement of RNA pol
Termination
 Occur at termination sites

 Occurs 1kbp from the site of 3’ end of RNA

 Nature of individual site is not known

 For a high level of transcription activators

bind to regulator elements called enhancers
that is located 100 – 200bp upstream the
initiation site