ASSIGMENT BIOLOGY_S1T3_ABDUL KAREM BIN

HALIM_SOALAN 3

QUESTION 3:

Subtopic 6.4 Gene Regulation and Expression–lac

operon

1.0: Definition and General Comments:

 Definition of operon
- Operon is operating units which can be defined as the cluster of genes located
together on the chromosomes and transcribed together. An operon required for the
transport and metabolism of lactose in E.coli and many other enteric bacteria. Gene
regulation of the lac operon was the first genetic regulatory mechanism to be
understood clearly, so it has become a foremost example of prokaryotic gene
regulation. Operon were first found as a method of gene expression control by
Francois Jacob and Jacques Monod in 1961.

 The process involved [transcription]

- Transcription is the process where a gene's DNA sequence is copied (transcribed)
into an RNA molecule. Transcription is a key step in using information from a gene to
make a protein. Gene expression is when a gene in DNA is "turned on," that is, used
to make the protein it specifies. Not all the genes in your body are turned on at the
same time, or in the same cells or parts of the body. For many genes, transcription is
the key on/off control point. For example, if a gene is not transcribed in a cell, it can't
be used to make a protein in that cell. As a result, the more a gene is transcribed, the
 more protein that will be made. Various factors that control how much a gene is
transcribed. For instance, how tightly the DNA of the gene is wound around its
supporting proteins to form chromatin can affect a gene's availability for transcription.
Proteins called transcription factors, however, play a particularly central role in
regulating transcription. These important proteins help determine which genes are
active in each cell of your body.

 The process involved [translation]

-Bacterial operons are polycistronic transcripts that are able to produce multiple
proteins from one mRNA transcript. In this case, when lactose is required as
a sugar source for the bacterium, the three genes of the lac operon can be
expressed and their subsequent proteins translated: lacZ, lacY, and lacA. The gene
product of lacZ is β-galactosidase which cleaves lactose, a disaccharide,
into glucose and galactose. lacY encodes Beta-galactoside permease, a membrane
protein which becomes embedded in the cytoplasmic membrane to enable
the cellular transport of lactose into the cell. Finally, lacA encodes Galactoside
acetyltransferase.

 Explanation on how each process connecting/relating to each

other

- A specific set of nucleotides along the template strand of DNA indicates where the
gene starts and RNA polymerase should attach and begin unravelling the double
helix. The section of DNA or the gene that is transcribed is known as the
‘transcription unit’. Rather than RNA polymerase which is moving along the DNA
strand, while the DNA moves through the RNA polymerase enzyme. As the template
strand moves through the enzyme, it is unravelled and RNA nucleotides are added to
the growing mRNA molecule. As the RNA molecule grows it is separated from the
template strand. The DNA template strand to reforms the bonds with its
complementary DNA and a double helix. In prokaryotic cells, such as bacteria, once
a specific sequence of nucleotides has been transcribed then transcription is
completed. This specific sequence of nucleotides is called the ‘terminator sequence’.
Once the terminator sequence is transcribed, RNA polymerase will detach from the
DNA template strand and releases the RNA molecule. No further modifications are
 required for the mRNA molecule and it is possible for translation to begin
immediately. Translation can begin in bacteria while transcription is still occurring.

 State what will happen to lac operon in the presence and absence
of lactose.

-When lactose is present, lac operon is switch on. Lactose molecules will enter E.coli
and then it will be converted into allolactose. Allolactose will binds to repressor
protein forming allolactose-repressor complex. It will change repressor protein
conformation. When it happened repressor protein unable to attach to the operator
gene.

When lactose is absent operon is switch off. The regulatory gene(lacI) is a codes for
repressor protein. Repressor protein can switch off the lac operon by binding to the
operator. Besides it also blocks attachment of RNA polymerase and make RNA
polymerase cannot bind to the promoter. Transcription of structural gene is also
blocked by repressor-operator complex. So lacZ, lacY and lacA genes are not
transcribed. Therefore, enzymes encodes β-galactosidase, permease and
transacetylase are not produced.

2.0: Lac operon and their functions

 State the components of lac operon

I. The regulatory gene (LacI)
II. Promoter
III. Operator
IV. Structural genes
V. Catabolyte activator protein (CAP)

 Explain promoter and its function

- Promoter defined as the binding site for RNA polymerase, the enzyme that performs
transcription. Besides promoter is a region of DNA where transcription of a gene is
initiated. Promoter are usually located near the transcription start sites of genes and
a vital component of expression vectors because they control the binding of RNA
 polymerase to DNA. RNA polymerase transcribes DNA to mRNA which is ultimately
translated into a functional protein.

 Explain operator function

- The operator is a binding site of repressor protein. This operator is a negative
regulatory site bound by the lac repressor protein. The operator overlaps with the
promoter, and when the lac repressor is bound, RNA polymerase cannot bind to the
promoter and start transcription. Operator genes contain the code necessary to begin
the process of transcribing the DNA message of one or more structural genes into
mRNA. Thus, structural genes are linked to an operator gene in a functional unit
called an operon.

 Explain structural genes and its function

 LacZ
-lacZ encodes β-galactosidase, which is an intracellular enzyme that hydrolyzes
the disaccharide lactose into glucose and galactose. LacZ act as encodes an
enzyme that splits lactose into monosaccharides (single-unit sugars) that can be
fed into glycolysis.

 LacY
- The LacY gene is a structural component of the lac operon (lactose operon)
system which enables bacteria, such as E.coli. The LacY gene codes for the
proteinBeta-galactoside permease. It is a transmembrane symporter found in the
cytoplasmic membrane that regulates the transport of lactose into the cell. LacY
similar as lacZ which it encodes a membrane-embedded transporter that helps
bring lactose into the cell.

 LacA
-lacA encodes β-galactoside transacetylase, which is an enzyme that transfers an
acetyl group from acetyl-CoA to β-galactosides.
 Diagram
3.0:Mechanism of lac operon in life

 Situation 1: A certain mutation in E.coli changes the lac operator

so that the active repressor cannot bind. How would this affect the
cell’s production of β-galactosidase? Explain.

- When mutation in E.coli changes the lac operator, the cell will continuously produce
β-galactosidase and the two of other enzymes for lactose utilization, even if it’s in the
absence of lactose, thus wasting cell resources.

 Situation 2: What will happen to the control of lac operon when an

individual drinks milk containing glucose? Explain.

- Low-level transcription of the lac operon occurs. The lac repressor is released from
the operator because the inducer (allolactose) is present. cAMP levels, however, are
low because glucose is present. Thus, CAP remains inactive and cannot bind to
DNA, so transcription only occurs at a low, leaky level.

 Explain the importance of lac operon

- The lac operon is found in E.coli and some other enteric bacteria. This operon
contains genes coding for proteins in charge of transporting lactose into the cytosol
and digesting it into glucose. This glucose is then used to make energy.The lac
operon of Escherichia coli is a classic model for studying gene regulation. This study
has uncovered features such as the environmental input logic controlling gene
expression, as well as gene expression bistability and hysteresis. Most lac operon
studies have focused on a few lab strains. The environmental dependence of lac
gene regulation in 20 natural isolates of E.coli and found a wide range of regulatory
responses. By transferring lac genes from natural isolate strains into a common
reference strain, regulation depends on both the lac genes themselves and on the
broader genetic background, indicating potential for still-greater regulatory diversity
following horizontal gene transfer. As a result there is substantial natural variation in
the regulation of the lac operon and indicate that this variation can be ecologically
meaningful.

4.0:conclusion
-The bacterium E. coli can grow in culture mediums containing a variety of energy
sources, including the sugar lactose. However, to use lactose, the bacterium must
first alter its metabolism. The bacterium must turn on several genes, found in
the lac operon, which are required for lactose metabolism.The lac operon is an
inducible system, meaning that the system is turned off until an inducer—lactose—
arrives on the scene. Other operons, such as the trp operon, work in the opposite
way: this system expresses genes in the operon until a repressor becomes activated
and turns the expression off.