ge class2
ge class2
GENE EXPRESSION
IN PROKARYOTES
& EUKARYOTES
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OBJECTIVES
To know and explain:
• Regulation of Bacterial Gene Expression
• Constitutive ( house keeping) vs. Controllable genes
• OPERON structure and its role in gene regulation
• Regulation of Eukaryotic Gene Expression at different levels:
• DNA methylation
• Histon modifications(Chromatin Remodeling)
• Increasing the number of gene copies (gene amplification)
• Changing the rate of initiation of transcription
• Alternate splicing
• mRNA stability
• Changing the rate of initiation of translation
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GENE EXPRESSION
Gene expression is the process by which the information
encoded in a gene is used to direct the assembly of a protein
molecule.
Gene expression is explored through a study of protein structure
and function, transcription and translation, differentiation and
stem cells.
It is the process by which information from a gene is used in the
synthesis of a functional gene product.
These products are often proteins, but in non-protein coding
genes such as ribosomal RNA (rRNA), transfer RNA (tRNA) or
small nuclear RNA (snRNA) genes, the product is a functional
RNA.
The process of gene expression is used by all known life -
eukaryotes (including multicellular organisms), prokaryotes
(bacteria and archaea)
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REGULATION OF GENE EXPRESSION
• Regulation of gene expression includes a wide range of
mechanisms that are used by cells to increase or decrease
the production of specific gene products (protein or RNA).
• Gene regulation is essential for viruses, prokaryotes and
eukaryotes as it increases the versatility and adaptability of
an organism by allowing the cell to express protein when
needed.
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• Although a functional gene product may be an RNA or a
protein, the majority of known mechanisms regulate protein
coding genes.
• Any step of the gene's expression may be modulated, from
DNA-RNA transcription to the post-translational modification
of a protein.
• The first discovered example of a gene regulation system was
the lac operon, discovered by Jacques Monod, in which
protein involved in lactose metabolism are expressed by
E.coli only in the presence of lactose and absence of glucose.
• Gene regulation drives the processes of cellular
differentiation and morphogenesis, leading to the creation of
different cell types in multicellular organisms where the
different types of cells may possess different gene expression
profile.
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GENES
Genes are subunits of DNA, the information
database of a cell that is contained inside the cell
nucleus.
This DNA carries the genetic blueprint that is used to
make all the proteins the cell needs.
Every gene contains a particular set of instructions
that code for a specific protein
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CLASSIFICATION OF GENE
WITH RESPECT TO THEIR
EXPRESSION
Constitutive ( house keeping) genes:
Are expressed at a fixed rate, irrespective to the cell
condition.
Their structure is simpler.
Controllable genes:
Are expressed only as needed. Their amount may
increase or decrease with respect to their basal level in
different condition.
Their structure is relatively complicated with some
response elements
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TYPES OF REGULATION OF GENE
POSITIVE REGULATION :
When the expression of genetic information is quantitatively
increased by the presence of specific regulatory element is
known as positive regulation.
Element modulating positive regulation is known as activator or
positive regulator.
NEGATIVE REGUATION:
When the expression of genetic information is diminished by
the presence of specific regulatory element is known as
negative regulation.
The element or molecule mediating the negative regulation is
said to be repressor.
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BIOLOGICAL SYSTEMS EXHIBITS 3
TYPES OF TEMPORAL RESPONSES:
Type A response:
• Increased extent of gene expression is continued in presence of
inducing signal.
• This is commonly observed in prokaryotes in response to
intracellular conc. of nutrient.
Type B response:
• Increased amount of gene expression Is transient even in
presence of regulatory signal.
• This is seen in commonly during development of organism.
Type C response:
• Increased gene expression that persists even after termination of
signal.
• It is seen in development of tissur or organ.
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PRINCIPLES OF GENE REGULATION
1) RNA polymerase binds to DNA at promoters.
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STEPS INVOLVING REGULATION OF
GENE EXPRESSION
Synthesis of the primary RNA transcript (transcription)
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STEP OF REGULATION IN PROKARYOTES
Transcriptional control
Translational control
Post translational control
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OPERON
In genetics, an operon is a functioning unit of genomic DNA
containing a cluster of genes under the control of a single
promoter.
Operons occur primarily in prokaryotes but also in some
eukaryotes.
Operons are related to regulons, stimulons and modulons.
An operon is made up of several structural genes arranged
under a common promoter and regulated by a common
operator.
It is defined as a set of adjacent structural genes, plus the
adjacent regulatory signals that affect transcription of the
structural genes.
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General structure of an operon
An operon is made up of 4 basic DNA components:
Promoter – a nucleotide sequence that enables a gene to be
transcribed. The promoter is recognized by RNA polymerase, which then
initiates transcription.
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GENERAL STRUCTURE OF AN
OPERON
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OPERON REGULATION
Operon regulation can be either negative or positive by induction or
repression.
Negative control involves the binding of a repressor to the
operator to prevent transcription.
In negative inducible operons, a regulatory repressor protein is normally
bound to the operator, which prevents the transcription of the genes on
the operon .
If an inducer molecule is present, it binds to the repressor and changes
its conformation so that it is unable to bind to the operator. This allows
for expression of the operon.
The lac operon is a negatively controlled inducible operon, where the
inducer molecule is allolactose.
In negative repressible operons, transcription of the operon normally
takes place.
The trp operon, involved in the synthesis of tryptophan (which itself acts
as the corepressor ), is a negatively controlled repressible operon.
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With positive control, an activator protein stimulates
transcription by binding to DNA.
In positive inducible operons, activator proteins are normally
unable to bind to the pertinent DNA.
When an inducer is bound by the activator protein, it undergoes a
change in conformation so that it can bind to the DNA and
activate transcription.
In positive repressible operons, the activator proteins are
normally bound to the pertinent DNA segment.
However, when an inhibitor is bound by the activator, it is
prevented from binding the DNA.
This stops activation and transcription of the system.
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THE ACTIVITY OF AN OPERON IN THE
PRESENCE OR THE ABSENCE OF REPRESSOR
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The lac operon
• The lac operon of the model bacterium Escherichia coli
was the first operon to be discovered and provides a
typical example of operon function.
• It consists of three adjacent structural genes, a promoter,
a terminator, and an operator.
• The lac operon is regulated by several factors including the
availability of glucose and lactose.
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Absence of lac operon:
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Presence of lac operon
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Activity of an lac operon in the
presence or absence of inducer
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TRYPTOPHAN OPERON
Discovered in 1953 by Jacques Monod and colleagues, the trp
operon in E. coli was the first repressible operon to be
discovered.
This operon contains five structural genes:
trp E,
trp D,
trp C,
trp B, and
trp A, which encodes tryptophan synthetase.
It also contains a promoter which binds to RNA polymerase and
an operator which blocks transcription when bound to the
protein synthesized by the repressor gene (trp R) that binds to
the operator
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Amount of trp operon affect transcription
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GENE REGULATION IN
EUKARYOTES
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Eukaryotic cells have a much larger genome
Eukaryotes have much greater cell specialization
Thus eukaryotic cells contain an enormous amount of DNA that
does not program the synthesis of RNA or protein
This requires complex organization
In eukaryotes expression of gene into proteins can be
controlled at various locations
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CHECK POINTS FOR GENE
EXPRESSION IN EUKARYOTES
• Synthesis of proteins is controlled right from the chromatin stage.
• Expression of gene is controlled at many steps during the process of
transcription and translation.
• Description of the control points is dealt in detail in the subsequent
slides.
SEVERAL STEPS OF EUKARYOTIC GENE
REGULATION
1.Transcriptional control.
2.RNAprocessing control.
3.RNA transport & localisation control.
4.Translation control.
5.mRNAdegradation control.
6.Protein activator control.
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1.CHROMATIN STRUCTURE
Two forms of chromatin
• Euchromatin – A lesser coiled transcriptionally
active region which can be easily accessed by the
RNA polymerases.
• Heterochromatin – A highly condensed
transcriptionally inactive region. The genes in this
region cannot be accessed by the RNA
polymerases for active transcription.
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MECHANISMS WHICH AFFECT THE
CHROMATIN STRUCTURE AND HENCE THE
EXPRESSION OF GENE ARE:
• Histone modifications – These modifications
make a region of gene either transcriptionally
active or inactive.
Acetylation
• ↑Acetylation ----↓ Condensation of DNA ----- ↑
Transcription of genes in that region
Ubiquitination
Ubiquitination of H2A – Transcriptional inactivation
Ubiquitination of H2B - Transcriptional activation
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METHYLATION
DNA methylation:is the addition or removal
of a methyl group predominantely where
cytosine bases occur consecutively.
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2.REGULATION OF TRANSCRIPTION
• Eukaryotes – There are two types of promoters which are:
i. Basal promoters
ii. Upstream promoters
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Transcriptional control:
… controlling when and how often a given gene is
Transcribed
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Enhancers
• Enhancers can be located upstream, downstream or
within the gene that is transcribed
• The binding of these enhancers with enhancer binding
proteins (transcription factors) increases the rate of
transcription of that gene to a greater extent.
• Promoters are capable of initiating lower levels of
transcription.
• Enhancers are responsible for the cell or tissue specific
transcription.
• Each enhancer has its own transcription factor that it
binds to.
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