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 - amanitin

RNA polyemerase II transcribe all protein coding genes and

some small nuclear RNA genes, present in nucleoplasm,
sensitive to 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 - 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