PROKARYOTIC

TRANSCRIPTION
Introduction

RNA molecules are synthesised by using a portion

of one strand of DNA as template in a
polymerization reaction.

Catalyzed by RNA polymerases

Process by which RNA molecules are initiated,

elongated and terminated Transcription
The Central Dogma

Transcription Translation
DNA RNA PROTEIN
Replication

Central dogma : Intracellular information flow from DNA to

mRNA to Protein.
3
 RNA Polymerases
Principal enzyme in transcription

Catalyzes polymerization of ribonucleotides as

directed by DNA template

Catalyses growth of RNA chains in 5' 3'

direction
Preformed primer is not required

Transcription initiates denovo at specific sites at

the beginning of gene

 RNA POLYEMERASE:
Complex enzyme made up of multiple polypeptide chain

6 subunits :
2 subunit binding of enzyme to promoter region

nucleotide addition

part of core enzyme , keeps the RNA bound to DNA template

part of enzyme, but no specific function

sigma factor, recognize promoter region binds to it. weakly

bound, which dissociates from enzyme during elongation
[ Core enzyme : free unit , ie., 2 Holoenzyme : the complete enzyme,
ie., 2 ]
One of the largest enzyme (mw 465,000Da) ,contact with many

DNA bases simultaneously

 Selection of site The Promoter
DNA sequence to which the RNA polymerase binds to initiate
transcription of gene Promoter

2 types of regions important in promoter function :

Pribnow Schaller box : (prokaryotes)
Located about 10 bp upstream from start of transcription

Goldberg Hogness box : (eukayotes)

Located 20-30 bp upstream from start of transcription
Also termed as TATA box.
Both are A-T rich regions.

Concensus sequences : basic sequence derived from a large set of

observed similar sequence
Steps : Transcription
Initiation of transcription

RNA chain elongation

Termination and release of newly synthesised RNA

 Initiation of Transcription
subunit binds specifically to sequence in both -35 and -10
promoter regions
subunit binds non specifically to DNA low affinity, initial binding ><
polymerase and promoter closed promoter complex (CPC), DNA is
not unwound
Polymerase unwinds 12-14 bases of DNA from about -12 to +2 to
form open promoter complex, ssDNA available as template for
transcription
RNA polymerase contains2 nucleotide binding sites - Iniation site &
elongation site
Initiation site binds to purine triphosphates , ATP & GTP
ATP first nucleotide in chain first DNA Base transcribed thymine
Transcription initiated by addition of NTPs
 Chain elongation
Polymerase + template continues synthesis of mRNAs
Polymerase unwinds DNA and adds up nucleotides upto 8 to growing
chain
Polymerase undergo confirmational changes & loses subunit
Along with unwinding of DNA , it rewinds the DNA behind
it,maintaining an unwound region (15bp)
8-12 bases of growing RNA chain remains bound to complementary
DNA
and subunit forms crab claw like structure grips DNA template
Elongation :
doesnt occur at constant rate
Reduction in rate Pause
Pausing follows sequences that forms hairpin in RNA
Continues till termination signal is reached
 Termination & release of newly sythesised RNA
Transcription stops and RNA released from polymerase & enzyme
dissociates from DNA template
Transcription of GC rich inverted repeat RNA segment, can form a
stable stemloop structure by base pairing
Self complementary structure disrupts association with DNA
Termination
H2 boding >< A and U is weaker than >< G and C
 TWO TYPES OF TERMINATION :

Rho() dependent

Rho independent
 Rho dependent termination
Rho - the termination factor protein

Transcription of some genes is terminated by specific termination protein :

Rho, an ATP dependent helicase

It moves along RNA transcript (>60 nt) , finds the "bubble", unwinds it and
releases RNA chain

Specific sequences - termination sites in DNA

inverted repeat, rich in G:C, which forms a stem-loop in RNA transcript

6-8 As in DNA coding for Us in transcript

Rho dependent termination
Rho independent termination
Termination occurs in absence of Rho protein

Terminator consist of:

A short inverted repeat followed by a stretch of A : T basepairs (about 8)

When polymerase transcribes inverted sequence , a stem and loop structure is

formed

mRNA moves away from DNA template as a A-U bp. it detaches and so RNA
polymerase cannot move forward by formation of such structre transcription
ends
Rho independent termination
Rho dependent termination Rho independent termination