TRANSCRIPTION

(General features of transcription, RNAP, Promoter,

Prokaryotic transcription initiation )
 TRANSCRIPTION
• Transcription means that the genetic information stored in double-
stranded DNA are copied in the form of a single-stranded RNA
molecule like mRNA, tRNA, rRNA
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Genes are composed of three sequence region

1.promoter region
2.RNA coding sequences
3.terminator region
STEPS IN TRANSCRIPTION
/
 Prokaryotic RNAP

• The polymerase is a multi-subunits holoenzyme

• The core RNAP has 5 subunit two α, β, β’,ω
• The first step in the making of RNAP is the dimerization of alpha
subunits. They will acts as a scaffold to bring two other subunits β, β’
• RNAP cannot start transcription on its own. Its require the help of
a factor known as σ factor.
• σ factor is a protein that allow the specific binding of RNAP to the
promoter sequence
• So the RNAP holoenzyme composed of 6 subunits two α, β, β’,ω
and σ
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.

• the σ subunit is crucial in the recognition of the promotor region in the

DNA to initiate the process of transcription
• However, the catalytic core comprises of α, β, β’ω
• σ associate with RNAP core at the time of transcription
• Once the initiation site is determined, the σ subunit dissociates from
the catalytic core

+ 
Sigma factor

RNAP Core Enzyme

RNAP Holoenzyme
.
 RNAP types and functions

RNAP I
• The enzyme is responsible for synthesizing rRNA
• It transcribes the gene in the nucleolus and synthesizes the rRNA in
the nucleus itself from where it is transported to cytoplasm via either
nuclear pore or by carrier proteins where it forms forms ribosomes
RNAP II
• The enzyme is responsible for synthesizing mRNA
• It transcribes proteins coding genes from the DNA into suitable
mRNAs that can further be processed to take part in translation
,

• The working of this enzyme directly influences the proteins that are to
be synthesized, if improper transcription of genes then would lead to
translation of faulty proteins, which can have a severe impact on our
body
RNAP III
• This enzyme is responsible for synthesizing transfer RNA
• tRNA is responsible for attaching aminoacids and making polypeptide
chain
Eukaryotic RNAP
• RNA polymerase is different from DNA polymerase in that it does
not require a primer and it also does not have proofreading activity.
• RNA polymerase doesn't have proofreading ability because RNA
molecules can tolerate some errors. These errors are removed during
splicing, and new copies of RNA are formed
Functions of RNAP
• α Binds regulatory proteins
• β- forms phosphodiester bond
• β’- Fixes RNAP to DNA template
• σ – recognize and binds to promoter region of DNA, initiate
transcription
Promoter

• A promoter, is a region of DNA upstream of a gene where relevant

proteins (such as RNAP and TFs) bind to initiate transcription of that
gene.
• They are regulatory, not actually transcribed, and control gene
expression in bacteria and eukaryotes.
• Promoter sequences located directly upstream or at the 5′ end of the
transcription initiation site
• In prokaryotes, only three types of promoter sequences are found
namely, -10 promoters, -35 promoter and upstream elements.
• In eukaryotes, there are many different promoter elements such as
TATA box, initiator elements, GC box, CAAT box, etc.
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Eukaryotic Gene Structure
.

Prokaryotic Gene Structure

PROMOTER STRUCTURE-Prokaryotes

• The transcription start point, which the first base

transcribed is termed the +1 site or the initiation site.
.
• Three main promoter elements
1.Upstream promoter element(UPE)
-located at -58,-37
-interact with the RNA polymerase (RNAP)
α-subunit and stabilizes it.
2. -35 element
-sequence:TTGACG
3. -10 element(pribnow-schaller box)
-Or pribnow box
-sequence:TATAAT
• Discriminator : is a spacing element, 5–8 base pairs (bp) long. It is
located between the -10 element and the transcription start site
2 type promoter

Strong promoter
• a promoter that increases the transcription of genes of interest and
enhances the production of various substances
• High rate of transcription
Weak promoter
• a promoter that directs relatively low amounts of expression.
• Low rate of transcription
• When the gene product is toxic to the cell, a weak promoter is used to
avoid cell death due to the over expression of the gene product
Stages in prokaryotic transcription

Initiation
Elongation
Termination
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INITIATION
sigma unit of RNA polymerase identifies
the promoter region on template DNA.
• RNA polymerase holoenzyme binds
loosely to DNA at first(-35,-10 region)
• It either binds initially at a promoter or
scans along the DNA until it finds one
• The complex with holoenzyme loosely
bounds at the promoter is called a closed promoter complex because
the DNA remains in closed double-stranded form.
 .

• The RNA pol then interacts with the

DNA and unwinds around 15 bases
long region at the initiation site to
create a transcription bubble,
collectively
called open-promoter complex.
• single-stranded DNA is exposed and
can be used as a template for transcription.
• The strand which is transcribed is known as the template strand
or antisense strand.
• Once the open complex is formed the enzyme starts to add rNTP’s on template DNA.
RNAP does not require primer for initiation

Abortive initiation
also known as abortive transcription
• Repetitive synthesis and release of short nascent RNAs(approximately
10 NTs) by RNA polymerase.
• The process take place till RNA polymerase-promoter initial
transcribing complex leave the promoter.
• The RNA pol escapes the promoter and initiates productive synthesis
of RNA, forming RNA polymerase-DNA elongation complex
.

• abortive initiation is a steady state assay that monitors functional

initiation complexes.
IMPORTANCE
• Dissociation of sigma factors and transcription factors.
• Promoter clearance.
• Proper orientation of DNA & RNA Polymerase complex for entering
elongation stage.