Regulation of Gene Expression 2016

Regulation of Gene Expression in Prokaryotes
Agus Limanto
Faculty of Medicine
UKRIDA
Outline
• Objectives
• Pathway of Gene expression
• Constitutive, Inducible, and Repressible Gene Expression
• Operon: A Coordinated Unit of Gene Expression
• Positive and Negative Control of Gene Expression
• Lactose Operon
• Tryptophan Operon
• Protein – DNA Interaction in Transcriptional Regulation
• Regulatory Mechanism in Viruses
Objectives
• After going through this lecture, student should be able to
understand:
• Constitutive and inducible proteins, enzyme repression and enzyme induction
• Fundamental structural design of the operon and its genetic components
• Roles of regulatory gene, repressor and corepressor, catabolite activator
protein (CAP), cyclic AMP and attenuator in case of lac operon and tryp
operon; regulatory mechanisms in viruses; and significance of protein-DNA
interaction in regulation of transcription
Pathway of Gene Expression
Constitutive, Inducible, and Repressible Gene Expression
• Constitutive genes
• Certain gene products – such as tRNA molecules, rRNA molecules, ribosomal proteins, RNA
polymerase subunits, and enzymes catalyzing metabolic processes - are essential components of
almost all living cells
• They are frequently referred as housekeeping genes
• Inducible proteins
• Synthesized only when the cellular need for them arises
• The process called as induction
• Catabolic pathway
• Classical example: β-galactosidase
Constitutive, Inducible, and Repressible Gene Expression
• Repressible proteins
• Classical example: tryptophan
• Synthesized only when the cell has to grow in a tryptophan-free medium
• Anabolic pathway
• The process called as repression
• Keypoints
• Transcriptional regulation is the most common
Operon: A Coordinated Unit of Gene Expression
• In bacteria, genes with related functions frequently occur in coordinately regulated units called operons
• Each operon contains a set of contiguous structural genes, a promoter (the binding site for RNA
polymerase), and an operator (the binding site for a regulatory protein called a repressor)
• When a repressor is bound to the operator, RNA polymerase can’t transcribe the structural genes in the
operon
• When the operator is free of repressor, RNA polymerase can transcribe the operon
Positive and Negative Control of Gene Expression
• Gene expression is controlled by both positive and negative regulatory mechanism
• In positive control mechanisms, the product of regulator gene, an activator, is required to turn on
the expression of the structural genes
• In negative control mechanisms, the product of a regulator gene, a repressor, is necessary to turn off
the expression of the structural genes
• Activators and repressors regulate gene expression by binding to sites adjacent to the promoters of
structural genes
• Whether or not the regulator proteins can bind to their binding sites depends on the presence or
absence of small effector molecules that form complexes with the regulator proteins
• The effector molecules are called inducers in inducible systems and co-repressors in repressible
systems
Lactose Operon in E. coli: Induction and Catabolite Repression
• E. coli lac operon is a negative inducible and catabolite repressible system

• The three structural genes in the lac operon are transcribed at high levels only in the presence
of lactose and the absence of glucose
• In the absence of lactose, the lac repressor binds to the lac operators and prevents
RNA polymerase from initiating transcription of the operon
• Catabolite repression keeps operons such as lac encoding enzymes involved in

carbohydrate catabolism from being induced in the presence of glucose
• The binding of CAP/cAMP complex to its binding site in the lac promoter bends the
DNA and makes it more accessible to RNA polymerase
• The lac repressor binds to two operators – either O1 and O2 or O1 and O3 –

simultaneously and bends the DNA into a hairpin or a loop, respectively
Lactose Operon in E. coli: Induction and Catabolite Repression
Tryptophan Operon in E. coli: Repression and Attenuation
• The E. coli trp operon is a negative repressible system

• Transcription of the five structural genes in the trp operon is repressed in the presence
of significant concentrations of tryptophan
• Operons such as trp that encode enzymes involved in amino acid biosynthetic pathways
often are controlled by a second regulatory mechanism called attenuation
• Attenuation occurs by the premature termination of transcription at a site in the mRNA

leader sequence when tryptophan is prevalent in the environment in which the bacteria are
growing
Protein – DNA Interaction in Transcriptional Regulation
• Role of protein – DNA interaction is of vital significance in regulating gene transcription
• Specific factors
• Regulatory proteins
• Specific factors
• Help RNA polymerase to recognize specific site where transcription begins
• Involved specific promoter recognition and binding
• Alteration in this protein will alter specificity of initiation
• Regulatory proteins
• Bind with DNA sequence present in the vicinity of the transcriptional initiation site
• Regulate the rate of transcription
• Allosteric proteins with binding and non-binding conformations
Regulatory Mechanism in Viruses
• There are two regulation

• Lysogenic pathway
• Virus integrates its DNA
into the host-cell
chromosome where it
replicates with the latter
• Lytic pathway
• Viral DNA replicates
independently till the host
cell is destroyed

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Regulation of Gene Expression in Prokaryotes

• E. coli lac operon is a negative inducible and catabolite repressible system

• Catabolite repression keeps operons such as lac encoding enzymes involved in

• The lac repressor binds to two operators – either O1 and O2 or O1 and O3 –

• The E. coli trp operon is a negative repressible system

• Attenuation occurs by the premature termination of transcription at a site in the mRNA

• There are two regulation

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