Location via proxy:   [ UP ]  
[Report a bug]   [Manage cookies]                
Scribd
Upload
11 views

Lecture 4 -4 (part 2)

Uploaded by

aze rty
Copyright
© © All Rights Reserved
Available Formats
Download as PDF, TXT or read online on Scribd
11 views

Lecture 4 -4 (part 2)

Uploaded by

aze rty
Copyright
© © All Rights Reserved
Available Formats
Download as PDF, TXT or read online on Scribd
You are on page 1/ 20

Regulation of Gene

Expression
came originally from the

Regulation of Gene Expression study of how E. coli


bacteria adapt to
changes in the
composition of their
growth medium.

Regulation of Gene Expression

! Genes, based on their activity, can be grouped as housekeeping genes and


others are classed as induced to express or express in stage specific or tissue
specific way.

! Genes turned on all the time (Constitutive) Housekeeping genes express all
the time under all normal conditions.

! Other genes can be regulated: – Turned On – Turned Off

! Regulation of gene expression allows cells to respond to changes in their


environment
General rule Gene expression is
regulated at the level of transcriptional
initiation
! An inducible gene is a gene that is expressed in the presence of a
substance (an inducer) in the environment. This substance can control the
expression of one or more genes (structural genes) involved in the
metabolism of that substance. For example, lactose induces the
expression of the lac genes that are involved in lactose metabolism. An
certain antibiotic may induce the expression of a gene that leads to
resistance to that antibiotic.

! Induction is common in metabolic pathways that result in the catabolism


of a substance and the inducer is normally the substrate for the pathway.

! Repressible gene is those in which the presence of a substance (a co-


repressor) in the environment turns off the expression of those genes
(structural genes) involved in the metabolism of that substance. e.g.,
Tryptophan represses the expression of the trp genes.

! Repression is common in metabolic pathways that result in the


biosynthesis of a substance and the co-repressor is normally the end
product of the pathway being regulated.
Two Major Types of Transcriptional Control
Positive Control: Genes under positive control only be expressed when an
activator protein is present.

Negative control: Genes under negative control are expressed unless a


repressor protein is present and blocks their transcription.
! Although induction and repression appear to be different processes,
both involve the activity of repressors, proteins that bind to DNA to
prevent initiation of transcription

! the levels of various repressors & activators of transcription depend


on the cellular environment, which thus determines which genes are
ON or OFF!

• Let’s see how this works in genes involved with metabolism in E. coli...

! The Lactose Operon: A Model for Inducible Gene Regulation in


Bacteria

! The Tryptophan Operon: A Model for Repressible gene Regulation


in Bacteria
The Tryptophan Operon
! Tryptophan: amino acid E. Coli can synthesize all
the 20 amino acids

Systematic name: 2-amino-3-(1H-indol-3-yl) propanoic acid

Molecular Formula: C11H11N2O2

Tryptophan operon:

series of structural genes all under the control of a Regulatory


Gene • Tryptophan operon is normally turned on
• Tryptophan operon is a repressible operon
Tryptophan synthesis regulate transcription of mRNA,
thus control enzyme synthesis.

chorismate is transformed into trp by 3 enzymes encoded by 5


genes organized in an operon structure:
anthranilate synthase (subunit I + subunit II) (genes trpE and
trpD) (anthranilate synthase+ phosphoribosyl transferase)
anthranilate isomerase (gene trpC)(PRA isomerase+InGP
synthase)
tryptophan synthase (a subunit + b subunit) (genes trpB and
trpA)
The Tryptophan Operon: A Model for Repressible
gene Regulation in Bacteria

! Structural genes: five genes trp E, trp D, trp C, trp B, and trp A, which
encode tryptophan synthetase.
! Regulatory region
• Trp R: which synthesizes a specific protein (repressor) which then causes
the transcription to be blocked.
• Promoter: where RNA polymerase binds to the operon
• Operator: a sequence that acts as an on/off switch for transcription
• Trp L: The sequence between the promoter and trpE gene has 2
interesting features
" A short coding sequence encoding a leader peptide of 14 amino acids
" An attenuator sequence encoding a GC rich transcript forming an hairpin
structure ending with a series of U residues (single stranded).
Two Mechanisms to Regulate the Expression
of trp Operon
1. Repression:

-Trp is high: inhibition of the trp operon by the repressor which inhibits the binding of RNA pol to the
promoter
-Binding of the repressor to the operator is regulated by level of trp; trp increases the affinity of the repressor
for the operator. Trp acts as a co-repressor (stimulates attachment of repressor to operator)
promoter operator leader attenuator trpE trpD

trp repressor

-Trp is low: repressor is inactive, operator is free and RNA pol binds to the promoter:
transcription of operator

promoter operator leader attenuator trpE trpD


Binding of the Repressor to the Operator of the trp
Operon

Repressor binds DNA as a dimer and binding is regulated by trp:


- trp increases the affinity of the repressor

Two molecules of repressor bind DNA and trp induces conformational changes
allowing binding to DNA; trp is an allosteric effector.
Attenuation of the trp Operon
One element of the trp operon is the leader sequence (L) that sequence between
the promoter and trpE gene has 2 interesting features:
1. A short coding sequence encoding a leader peptide of 14 amino acids
2. An attenuator sequence encoding a GC rich transcript forming an hairpin
structure ending with a series of U residues (single stranded).
Mechanisms to Regulate the Expression of trp
Operon
! Attenuation:
Mechanism by which RNA pol can terminate transcription of the trp operon
before the structural genes. The leader region (160 nts) has a sequence capable
of forming hairpin loops.

! This transcript includes four short sequences specified 1-4, each of which is
partially complementary to the next one. Thus, three specific secondary
structures(hairpins) can form: 1-2, 2-3 or 3-4 of the leader mRNA.

! If region 3 is hybridized with region 4, the hairpin structure will be close


enough to a series of U residues single stranded: this is a Stop signal for
RNA pol.
The Attenuator of the Trp Operon
• The hybridization of sequences 1 and 2 to form the 1-2 structure is rare because
the RNA Polymerase waits for a ribosome to attach before continuing translation
past sequence 1.
• The formation of a hairpin loop between sequences 2-3 prevents the formation of
hairpin loops between both 1-2 and 3-4.
• leader peptide contains two consecutive codons for trp
• If the ribosome tries to translate this peptide while tryptophan levels in
the cell are low, it will stall at either of the two trp codons. While it is
stalled, the ribosome physically shields sequence 1 of the transcript, thus
preventing it from forming the 1-2 secondary structure. Sequence 2 is
then free to hybridize with sequence 3 to form the 2-3 structure, which
then prevents the formation of the 3-4 termination hairpin, thus the 2-3
structure is called anti-termination hairpin. RNA polymerase is free to
continue transcribing the full operon.
! If tryptophan levels in the cell are high, the ribosome will translate the
full leader peptide without interruption and will only stall during
translation termination at the stop codon. At this point the ribosome
physically shields both sequences 1 and 2. Sequences 3 and 4 are thus
free to form the 3-4 structure which terminates transcription.
Resume: Regulation
of Expression of the trp
Operon

Initiation of transcription of the trp operon takes place at


the level of the promoter.
In absence of tryptophan, RNA polymerase is able to
initiate transcription.
In presence of tryptophan, RNA polymerase cannot
initiate transcription because the operator site is
occupied by the repressor.

When the repressor is not present on the operator, RNA


polymerase initiates transcription and passes through the
attenuator. Translation starts.

When levels of tryptophan are low, ribosomes pause in


region 1, no terminator structure is formed (regions 2 and 3). Transcription continues.
In the presence of higher levels of tryptophan, the
secondary structure of a terminator is formed
(regions 3 and 4) and transcription terminates
prematurely.

Pearson Education 2009

You might also like