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Name: Jason Lloyd Fantonalgo Score:

Nixon Rauff Fowler Date: November 30, 2023
Manuel Castones Legarde III

EXERCISE NO. 4

The Operon Systems

OBJECTIVES

The students should be able to:

1. build genetic regulatory models for prokaryotes showing the various

components and their interactions;
2. differentiate an inducible and a repressible system of gene regulation in
prokaryotes.

MATERIALS

Two sets of colored pieces of cardboard per group

Scissors

PROCEDURE

1. Each group must construct a model for the lactose operon (inducible system) and the
tryptophan operon (repressible system). Drawings must be made of the two operons
in the turned on and turned off conditions.

2. Using the set of colored pieces of cardboard, cut out structural components of the
regulatory systems to fit in a jigsaw puzzle. Each component should vary in either
color or shape.

3. Cut out the other functional components (e.g. enzymes or other proteins, repressors,
co-repressors, inducers, signaling agents, etc.). Show how these other components
will fit each other and with the structural components.

4. As soon as the models are completed, demonstrate how the models will work in the
regulation of gene expression.
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SKETCHES

Lactose Operon in the Turned-off Condition

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Tryptophan Operon in the Turned-on Condition

Tryptophan Operon in the Turned-Off Condition

QUESTIONS

1. Genetic expression in general can be regulated at various stages. Identify specific

processes that control gene expression at the:

a. transcription level
- Many different mechanisms control gene regulation. One of the most
important is transcription factors. Transcription factors are proteins that
bind to specific DNA sequences and regulate the rate of transcription,
which is the process of copying DNA into RNA. Some transcription factors
activate transcription, while others repress it. Another important
mechanism of gene regulation is epigenetic modifications. Epigenetic
modifications are changes to DNA that do not change the DNA sequence
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itself, but can affect gene expression. For example, DNA methylation can
silence genes by preventing transcription factors from binding to DNA.
Chromatin remodeling is another important mechanism of gene regulation.
Chromatin is the complex of DNA and proteins that make up
chromosomes. Chromatin remodeling involves changing the structure of
chromatin, which can affect the accessibility of DNA to transcription
factors.

b. post-transcription level
- Post-transcriptional gene regulation involves essential processes. RNA
splicing, a fundamental mechanism, removes introns and joins exons,
creating mature mRNA ready for translation; RNA stability controls
mRNA lifespan before degradation; RNA export transports mRNA from
the nucleus to the cytoplasm for protein synthesis. These processes
collectively fine-tune gene expression beyond transcription, ensuring
precise cellular responses.

c. translation level
- Gene expression regulation during translation involves key components.
Translation initiation factors, specialized proteins, kickstart translation by
binding mRNA and ribosomes. MicroRNAs, small RNA molecules, can
hinder translation by binding to mRNA. Codon bias, which is the
preference for specific codons, influences translation speed by affecting
tRNA availability. These elements work together to finely control gene
expression during the translation process, adapting protein synthesis to the
cell's requirements.

2. Do you expect operons to occur extensively in eukaryotes? Explain.

No, I do not expect operons to occur extensively in eukaryotes. Operons are a

hallmark of prokaryotic genomes, where they allow concerted expression of functionally
related genes as single polycistronic transcripts. In contrast, eukaryotic genes are
generally considered monocistronic, each with its promoter at the 5' end and a
transcription terminator at the 3' end. When a segment of replicating DNA comprises
multiple cistrons, it is called "polycistronic," whereas if the DNA segment contains only a
single cistron or gene, it is referred to as "monocistronic."
Furthermore, operons are groups of genes that are transcribed together into a
single mRNA molecule. This allows for the coordinated expression of genes that are
involved in the same biological process. Operons are common in prokaryotes, but they are
rare in eukaryotes. One reason for this is that eukaryotic genes are often interrupted by
introns. Introns are non-coding sequences that must be removed before the mRNA can be
translated. This process, called RNA splicing, is not possible with polycistronic mRNA
molecules, which are produced by operons. Another reason is that multiple factors,
including transcription factors, enhancers, and silencers often regulate eukaryotic genes.
These factors can act independently or together to control gene expression. It would be
difficult to coordinate the regulation of multiple genes in an operon in such a complex
system.

3. When lactose is added to a growing culture of E. coli, the cells begin to make enzymes
necessary for lactose utilization. In contrast, when tryptophan is added to the culture
medium, the synthesis of enzymes of the tryptophan pathway ceases. Contrast the two
systems explaining why they act in reverse directions.

The lactose and tryptophan operons function differently because they serve distinct
purposes in E. coli. The lactose operon activates genes needed to break down lactose
sugar, a task required only when lactose is present in the environment. Thus, it is an
inducible operon, turning on in response to lactose's presence, saving energy by not
producing unnecessary enzymes. Conversely, the tryptophan operon is repressible,
shutting off genes that produce the amino acid tryptophan, a constant requirement. This
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mechanism prevents E. coli from overproducing tryptophan, which can be harmful. In

essence, these operons adapt their regulation to suit the specific needs of the bacterium in
varying environmental conditions.
In simpler terms, the lactose operon turns on genes when lactose sugar is present,
and the tryptophan operon turns off genes when tryptophan is present. This is because E.
coli only needs to break down lactose sugar when it is present, but it always needs to be
able to make tryptophan.

4. Attardi et al (1983) found out that lactose operon mRNA synthesis in E. coli increased
with the addition of an inducer. Do the inducers act at the transcription or translation
level? Explain.

Inducers are molecules that turn on genes. In the case of the lactose operon in E.
coli, the inducer is lactose itself. In the absence of lactose, the lac repressor protein binds
to the lac operator, which is a DNA sequence that blocks the RNA polymerase from
binding to the promoter and initiating transcription of the lactose operon. This means that
the genes in the lactose operon are not turned on. When lactose is present, it binds to the
lac repressor protein and causes it to change shape. This prevents the lac repressor from
binding to the lac operator, which allows the RNA polymerase to bind to the promoter
and initiate transcription of the lactose operon. This means that the genes in the lactose
operon are turned on and the enzymes needed to metabolize lactose are produced.
Therefore, inducers act at the transcription level to increase lactose operon mRNA
synthesis in E. coli by preventing the lac repressor protein from binding to the lac
operator.

REFERENCES:

BYJU’S Tuition Centre. (n.d.). Lac Operon - Regulation of Gene Expression. BYJU’S. Retrieved
October 13, 2023, from
https://byjus.com/biology/lac-operon-regulation-gene-expression/#:~:text=In%20lac%20o
peron%2C%20lactose%20acts,allows%20transcription%20of%20the%20operon.

Hornstein, N., & Grohmann, K. (2016, December). Beneath the surface. Inference. Retrieved
October 13, 2023, from https://inference-review.com/letter/beneath-the-surface

Phillips, T. (2008) Regulation of transcription and gene expression in eukaryotes. Nature

Education 1(1):199

Review of the Lac Operon & TRP Operon. (n.d.). [Video]. Pearson+. Retrieved October 13, 2023,
from
https://www.pearson.com/channels/microbiology/learn/jason/ch-16-central-dogma-gene-r
egulation/review-of-the-lac-operon-trp-operon-Bio-1