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Gene transcription in prokaryotes
By,
Dr. Komal Acharya
Gene expression in prokaryotic cell
RNA (Ribonucleic acid)
Types of RNA
• It is a segment of the DNA that is to be transcribed
into RNA.
• It has transcription start site and transcription termination site.
• Upstream region: sequence prior to the start site
• Downstream region: sequence after the start site.
• Template strand: a strand used as template
• Nontemplate/coding strand is complementary to template
strand
 Transcription is the first step pf the gene expression.
 It is a process of formation of the RNA transcript from the DNA.
 What does a cell require for transcription?
1. Transcription proteins/enzymes
2. Ribonucleotides
3. Template strand
Transcription unit
Promoter
• It is a DNA sequence where the RNA polymerase binds first.
• It provides signal for the initiation of transcription process.
• It has two consensus sequences:
1. – 10 consensus sequence / Pribnow box : TATAAT sequence
2. – 35 consensus sequence : TTGACA
• It catalyses the synthesis of RNA from DNA.
• The RNA polymerase core enzyme is made upof α, β and β’, ω subunits
• that attached to the σ- factor and together they make holoenzyme that carry out the
transcription process.
RNA polymerase
Subunit Function
α- subunit Assembly of core enzyme, promoter recognition, interaction with
regulatory factors
β and β’ Together they make catalytic centre. Β involved in chain elongation
ω Assembly and stabilization of RNA polymerase holoenzyme
σ Promoter recgnition
Transcription is divided in three steps
Initiation
Elongation
Termination
RNA polymerase holoenzyme binds at the promoter and form
a close binary complex
After that, melting of the short DNA strand bound to the
enzyme occurs and as a result, open binary complex is
formed.
Unwinding of 12-14 bp occurs which results in formation of
transcription bubble.
Some new nucleotides are incorporated followed by
phosphodiester bond formation. So, the ternary complex is
generated that contains DNA-RNA-enzyme.
Initiation
After that, σ – factor is released and the core enzyme remained
at the promoter. The enzyme leaves promoter region after
addition of 8-9 nucleotides.
 During elongation, the transcription bubble moves on further.
 Temporary DNA-RNA hybrid duplex is formed within the transcription bubble.
 The RNA polymerase moves along the DNA helix and keep on unwinding it.
 The new nucleotides are added at the 3’ end of the growing RNA chain.
 As the elongation continues, two strands behind the transcription bubble resume
their double helical structure.
 During whole process, as the RNA polymerase moves forward the DNA, it generates
positive supercoiling ahead and negative supercoiling behind the transcription
bubble. So, gyrase and topoisomerase I participate to release these supercoils.
Elongation
Termination
Rho-dependent termination Rho-independent/intrinsic termination
• Rho-protein is needed.
• It binds to the growing RNA chain at rut-site and travel along the RNA in
5’ to 3’ direction till it reaches to DNA-RNA hybrid.
• When RNA polymerase reaches at the terminator it catches up with the
Rho-protein and release the RNA.
• Intrinsic terminator has GC rich inverted repeats. So, the RNA transcript will
have abundance of GC base pairs that results in formation of hairpin loop like
structure.
• This loop in the transcript is followed by U-rich region where the RNA-
polymerase stops for some time and the poly U region is too weak to hold
the DNA:RNA duplex. So, the RNA is released followed by dissociation of the
enzyme.

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transcription.pptx

  • 1. Gene transcription in prokaryotes By, Dr. Komal Acharya
  • 2. Gene expression in prokaryotic cell RNA (Ribonucleic acid) Types of RNA
  • 3. • It is a segment of the DNA that is to be transcribed into RNA. • It has transcription start site and transcription termination site. • Upstream region: sequence prior to the start site • Downstream region: sequence after the start site. • Template strand: a strand used as template • Nontemplate/coding strand is complementary to template strand  Transcription is the first step pf the gene expression.  It is a process of formation of the RNA transcript from the DNA.  What does a cell require for transcription? 1. Transcription proteins/enzymes 2. Ribonucleotides 3. Template strand Transcription unit
  • 4. Promoter • It is a DNA sequence where the RNA polymerase binds first. • It provides signal for the initiation of transcription process. • It has two consensus sequences: 1. – 10 consensus sequence / Pribnow box : TATAAT sequence 2. – 35 consensus sequence : TTGACA • It catalyses the synthesis of RNA from DNA. • The RNA polymerase core enzyme is made upof α, β and β’, ω subunits • that attached to the σ- factor and together they make holoenzyme that carry out the transcription process. RNA polymerase Subunit Function α- subunit Assembly of core enzyme, promoter recognition, interaction with regulatory factors β and β’ Together they make catalytic centre. Β involved in chain elongation ω Assembly and stabilization of RNA polymerase holoenzyme σ Promoter recgnition
  • 5. Transcription is divided in three steps Initiation Elongation Termination RNA polymerase holoenzyme binds at the promoter and form a close binary complex After that, melting of the short DNA strand bound to the enzyme occurs and as a result, open binary complex is formed. Unwinding of 12-14 bp occurs which results in formation of transcription bubble. Some new nucleotides are incorporated followed by phosphodiester bond formation. So, the ternary complex is generated that contains DNA-RNA-enzyme. Initiation After that, σ – factor is released and the core enzyme remained at the promoter. The enzyme leaves promoter region after addition of 8-9 nucleotides.
  • 6.  During elongation, the transcription bubble moves on further.  Temporary DNA-RNA hybrid duplex is formed within the transcription bubble.  The RNA polymerase moves along the DNA helix and keep on unwinding it.  The new nucleotides are added at the 3’ end of the growing RNA chain.  As the elongation continues, two strands behind the transcription bubble resume their double helical structure.  During whole process, as the RNA polymerase moves forward the DNA, it generates positive supercoiling ahead and negative supercoiling behind the transcription bubble. So, gyrase and topoisomerase I participate to release these supercoils. Elongation
  • 7. Termination Rho-dependent termination Rho-independent/intrinsic termination • Rho-protein is needed. • It binds to the growing RNA chain at rut-site and travel along the RNA in 5’ to 3’ direction till it reaches to DNA-RNA hybrid. • When RNA polymerase reaches at the terminator it catches up with the Rho-protein and release the RNA. • Intrinsic terminator has GC rich inverted repeats. So, the RNA transcript will have abundance of GC base pairs that results in formation of hairpin loop like structure. • This loop in the transcript is followed by U-rich region where the RNA- polymerase stops for some time and the poly U region is too weak to hold the DNA:RNA duplex. So, the RNA is released followed by dissociation of the enzyme.