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Information Readout: Transcription
Information Readout: Transcription
Information Readout: Transcription
READOUT:
TRANSCRIPTION
TRANSCRIPTION
Mechanistically similar DNA replication
in terms of substrates and template-
directed growth
2 major difference:
Only 1 DNA template strand is transcribed
for a particular gene
Only a small fraction of the entire genetic
potential of an organism is relaized in one
cell.
EXISTENCE OF mRNA
Until about 1960 it was thought that
rRNA represented the set of templates
fro protein synthesis
Jacob and Monod predicted the
existence of mRNA by analyzing E.coli
mutants that are altered in the control of
lactose metabolism
Operon model
T2 and T4 barcteriophage
CHARACTERISTICS OF
mRNA
There is a high rate of mRNA synthesis
followed by rapid degradation
Because of the rapid synthesis and
degradation, mRNA accumulates rapidly but
not to high steady state levels
mRNA, being a copy of 2 or more
contiguous genes, is large and
heterogenous
mRNA nucleotide sequence is identical to
one of the template strand of DNA
RNA POLYMERASE
Polynucleotide phosphorylase was initially
thought to be the RNA-synthesizing enzyme
Ultimately it turned out that it participates in
the degradation of bacterial mRNA
In prokaryotes, a single RNA polymerase
catalyzes the synthesis of all 3 RNA classes
In eukaryotes, 3 classes of polymerase
catalyze the different types of RNA
RNA POLYMERASE
The 3 classes of RNA Polymerase differ
in their sensitivity to inhibition by α-
amanitin
2 more inhibitors were used to confirm
other characteristics of transcription
Cordycepin
○ Transcription chain terminator
Actinomycin D
○ Intercalating agent
RNA POLYMERASE
KINETICS DNA POLYMERASE RNA POLYMERASE
ω 11,000 1 unknown
RNA POLYMERASE STRUCTURE
INITIATION OF TRANSCRIPTION
ELONGATION: Incorporation of
Ribonucleotides
PROMOTER RECOGNITION
The most frequently transcribed genes
initiate transcription about once every
10sec,whereas some genes are
transcribed as infrequently as once per
generation (30-60 min)
Rate –limiting step
Each gene transcribed in E.coli shared a
short adenine and thymine rich
sequence (CONSENSUS SEQUENCE)
CONSERVED SEQUENCES IN
PROMOTERS
TERMINATION
In bacteria, 2 distinct types of
termination events exists:
Factor –Independent
Factor – Dependent
FACTOR-INDEPENDENT
2 structural
features:
The symmetrical
GC-rich segments
that in the trasncript
have the potential
to form a stem-loop
structure
A downstream run
of four to eight A
residues
FACTOR DEPENDENT
TERMINATION
Less frequent
Rho protein is a
hexamer composed of
identical subunits
It is an RNA –DNA
helicase which
contains nucleotide
triphosphatase acivity
that is activated by
binding to
polynucleotides
REGULATION OF TRANSCRIPTION