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THE MECHANISM OF
TRANSCRIPTION IN BACTERIA
(CHAPTER 6)
6-1
DR MOHD NAZMI ABD MANAP
The flow of genetic information according to the central dogma of molecular
biology. 6-2
Basic Principles of
Transcription and Translation
• Transcription • During transcription
– Is the synthesis of – The gene determines the
RNA under the sequence of bases along the
direction of DNA length of an mRNA molecule
– Produces messenger DNA Gene 2
RNA (mRNA) molecule
Gene 1
• Translation Gene 3
– Is the actual
synthesis of a DNA strand 3
(template) A C C A A A C C G A G T
5
polypeptide, which
occurs under the TRANSCRIPTION
ribosomes 6-3
Protein Trp Phe Gly Ser
Amino acid
• In prokaryotes
– Transcription and translation occur
simultaneously
DNA
TRANSCRIPTION
mRNA
Ribosome
TRANSLATION
Polypeptide
Nuclear
envelope
TRANSCRIPTION DNA
Pre-mRNA
RNA PROCESSING
mRNA
Ribosome
TRANSLATION
(b) Eukaryotic cell. The nucleus provides a separate
Polypeptide compartment for transcription. The original RNA
transcript, called pre-mRNA, is processed in various
ways before leaving the nucleus as mRNA.
6-5
OBJECTIVE
• Topics :
Classification of RNA Molecules
Transcription Phases
Transcription vs Replication
RNA Polymerase Structure
Promoters
Transcription Initiation
Elongation
Termination of Transcription
6-6
CLASSIFICATION OF RNA MOLECULES
mRNA – messenger RNA : rRNA – ribosomal RNA :
-bring information from the DNA -80% of total RNA
-Translated into protein -Component of ribosome
-Not stable -Involve in protein synthesis
-stable
RNA molecules
Transcription
occurs in
three
phases:
1. Initiation
2. Elongation
3. Termination
6-9
Synthesis of an RNA Transcript
Promoter
Transcription unit
• The stages of
5 3
3 5
DNA
Start point
RNA polymerase 1 Initiation. After RNA polymerase binds to
transcription the promoter, the DNA strands unwind, and
the polymerase initiates RNA synthesis at the
start point on the template strand.
are 5
3
3
5
DNA transcript
2 Elongation. The polymerase moves downstream, unwinding the
DNA and elongating the RNA transcript 5 3 . In the wake of
– Initiation Rewound transcription, the DNA strands re-form a double helix.
RNA
– Elongation 5
3
5
3
3
5
RNA
– Termination transcript
3 Termination. Eventually, the RNA
transcript is released, and the
polymerase detaches from the DNA.
5 3
3 5
5 3
6-18
Structure of RNA polymerase
6-19
RNA POLYMERASE STRUCTURE
& FUNCTION
holoenzyme - 2a, 1b, 1 b’, & 1s.
6-24
UP ELEMENT
• Some very strong promoters have an
additional element (UP element) upstream
the core promoter – makes these
promoters even more attractive to RNA
polymerase - The rrnB P1 gene Promoter
6-25
TRANSCRIPTION INITIATION
• Transcription initiation was assumed to
end as RNA polymerase formed 1st
phosphodiester bond
6-32
E. coli s70
• Four regions
of high
sequence
similarity are
indicated
• Specific areas
that recognize
the core
promoter
elements, -10
box and –35
box are notes 6-33
Region 1
6-34
Region 2
• This region is the most highly conserved
of the four
• appears to be a-helix
6-35
Regions 3 and 4
• Region 3 is involved in both core and
DNA binding
• Nucleotides are T A G G T T
added sequentially, 5
Direction of transcription
Template
one after another in (“downstream”)
strand of DNA
6-39
FUNCTION OF THE CORE
POLYMERASE
• Core polymerase contains the RNA
synthesizing machinery
6-43
INVERTED REPEATS AND HAIRPINS
• A transcript of this
sequence is self-
complementary
– Bases can pair up to
form a hairpin as seen
in the lower panel 6-44
Rho-Independent termination
• The polymerase paused
with the formation of
hairpin
6-46
MECHANISM OF RHO
• No string of T’s in the
r-dependent
terminator, just
inverted repeat to
hairpin
• Binding to the growing
transcript, r follows
the RNA polymerase
• It catches the
polymerase as it
pauses at the hairpin
• Releases transcript
from the DNA-
polymerase complex
by unwinding the RNA-
DNA hybrid 6-47
SUMMARY : TRANSCRIPTION
• Is the generation of an mRNA copy of the
genetic information coded in the DNA
The extension of RNA chains takes place within the transcription bubble
(a locally unwound segment of DNA = 17-18 bp)
RNA pol continuosly unwinds the dsDNA ahead of the polymerization site &
rewinds the cDNA strand behind the polymerization site as it moves along the
double helix
The RNA chain is displaced from the DNA template strand as RNA pol moves
along the DNA mol – the region of transient base-pairing between the
growing chain & the DNA template strand is very short, only 3 bp
(hybridization of RNA & DNA template) 6-50
TRANSCRIPTION : TERMINATION
rho-independent terminator =
inverted repeat & T rich rho-dependent terminator –
form hairpin structure inverted repeat