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BML 1211 Biotechnology for Medical lab science - Lecture V

Biotechnology for Medical lab science - Lecture V

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BML 1211 Biotechnology for Medical lab science - Lecture V

Biotechnology for Medical lab science - Lecture V

Uploaded by

minute “minutemike” mike
Copyright
© © All Rights Reserved
We take content rights seriously. If you suspect this is your content, claim it here.
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BML 1211 BIOTECHNOLOGY FOR MEDICAL LAB SCIENCE – Lecture V

5.0 GENE STRUCTURE AND ORGANIZATION

A simple and universal definition of a gene is any DNA sequence that is transcribed into
an RNA molecule. Genes include DNA sequences that encode proteins, as well as
sequences that encode rRNA, tRNA, snRNA, and other types of RNA.

 Structural genes encode proteins that are used in metabolism or biosynthesis or


that play a structural role in the cell.

 Regulatory genes are genes whose products, either RNA or proteins, interact with
other DNA sequences and affect the transcription or translation of those sequences.
In many cases, the products of regulatory genes are DNA-binding proteins.

5.1 GENE REGULATION

Bacteria and eukaryotes use regulatory genes to control the expression of many of their
structural proteins. A gene does not only comprise the coding sequence, but also other
sequences required for the control of the expression of that coding sequence. These
DNA sequences are not transcribed but play a role in controlling gene expression.

 Transcriptional control elements – they include promoter and terminator.


The promoter marks the point at which transcription of the gene should start. It is
where the enzyme RNA polymerase binds to begin transcription. The terminator
marks the end of the gene where transcription should stop.

 Regulatory elements - These are short DNA sequences that affect the ability of
RNA polymerase to bind to the promoter. There are two types of regulatory elements
in eukaryotes. Enhancers that stimulate transcription and silencers that inhibit
transcription. In prokaryotes regulatory elements are known as operators.

Gene regulation in both bacterial and eukaryotic cells, takes place through the action of
regulatory proteins produced by regulatory genes that recognize and bind to regulatory
elements. Regulation that stimulates gene expression is termed positive control,
while regulation that inhibits gene expression is termed negative control.
A regulator gene helps to control the transcription of the structural genes, however it’s
not considered to be part of the gene. The regulator gene has its own promoter and is
transcribed into a short mRNA, which is translated into a small protein. This regulator
protein binds to the operator and affects transcription. Regulatory proteins are either
repressors or activators.
 Repressor - a regulatory protein that binds and inhibits transcription
 Activator - a regulatory protein that increases the transcription rate.

Together with regulatory proteins, small effector molecules often play a critical role in
transcriptional regulation. These small effector molecules exert their effects by binding
to the regulatory protein. The binding of the effector molecule causes a conformational
change in the regulatory protein affecting whether or not it can bind to the DNA.

An inducer is an effector molecule that causes transcription to increase by;


 binding to a repressor protein and preventing it from binding to the DNA, or
 binding to an activator protein and cause it to bind to the DNA.
Genes that are regulated in this manner are called inducible genes.

A corepressor is an effector molecule that inhibits transcription by binding to a


repressor and causing it to bind to the DNA. An inhibitor is an effector molecule that
binds to an activator and prevents it from binding to DNA. Both corepressors and
inhibitors act to reduce the rate of transcription. Therefore, the genes they regulate are
termed repressible genes.

5.2 BACTERIAL GENES

Bacterial genes that code for functionally related proteins, for example proteins that act
together to catalyze a metabolic pathway, are often arranged sequentially at specific
locations on the chromosome or plasmid. Such DNA sequences are known as operons.
The mRNA synthesized by the transcription of an operon is polycistronic, i.e., it
contains the information sequences of several genes. The information sequences are
separated by intercistronic regions. Each cistron has its own start and stop codon in
the mRNA. Operons are usually under the control of a single sequence known as
promoter. RNA polymerase binds to the promoter and then moves downstream,
transcribing the structural genes. Since the promoter initiates transcription, it is
therefore the point of control for the expression of the genes. Operons and polycistronic
mRNAs are common in bacteria but not in eukaryotes.

 Inducible operons are those in which transcription is always off. Something


must happen to induce transcription, or turn it on.
 Repressible operons are those in which transcription is always on. Something
must happen to repress transcription, or turn it off.

5.2.1 The lac operon

Lactose is a major carbohydrate found in milk. To metabolize lactose as an energy


source, E. coli produces enzymes β-galactosidase, transacetylase. The lac operon is an
example of a negative inducible operon. When lactose is absent from the medium in
which E. coli grows, few molecules of these enzymes are produced. If lactose is added to
the medium and glucose is absent, the rate of synthesis of all lac operon proteins
simultaneously increases. When lactose is absent, a repressor is bound to the operator,
blocking RNA polymerase from binding to the promoter and transcription is prevented.
5.3 EUKARYOTIC GENES

Eukaryotic genes are not organized into operons and are rarely transcribed together into
a single mRNA molecule. Instead, each structural gene typically has its own promoter
and is transcribed separately. Most eukaryotic genes, particularly those found in
multicellular species, are regulated by many factors. The term transcription factor is
broadly used to describe proteins that influence the ability of RNA polymerase to
transcribe a given gene. The two categories of eukaryotic transcription factors are.

 General transcription factors, which are required for the binding of RNA
polymerase to the promoter and its progression.

 Regulatory transcription factors serve to regulate the rate of transcription


of target genes. They recognize and bind to regulatory elements that are
located near the promoter. These DNA sequences are similar to the operator sites
found in bacterial. When a regulatory transcription factor binds to a regulatory
element, it affects the transcription of an associated gene. For example, the
binding of regulatory transcription factors may enhance the rate of transcription.
Such a transcription factor is termed an activator, and the sequence it binds to
is called an enhancer. Alternatively, regulatory transcription factors may act as
repressors by binding to elements called silencers and prevent transcription.
Many eukaryotic genes contain coding regions called exons and noncoding regions
called intervening sequences or introns. All the introns and the exons are initially
transcribed into RNA but, after transcription, the introns are removed by splicing and
the exons are joined to yield the mature RNA. Introns are rare in bacterial genes.

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