Running Head: TRP OPERON MUTATIONS ASSIGNMENT 1

How does gene regulation involving the trp repressor protein differ from what you

observed with the lac repressor in the previous lesson?

The trp operon is found in E. coli bacteria and is a group of genes that encode

biosynthetic enzymes for the amino acid tryptophan (Khan, n.d). When tryptophan levels are

low, the trp operon is expressed meaning it is “turned on”, however when tryptophan levels are

high, the trp operon is repressed meaning it is “turned off” (Khan, n.d). Overall the trp operon is

regulated by the trp repressor. Once bound to tryptophan, the trp repressor blocks expression of

the operon. Tryptophan is also regulated by a mechanism based on coupling of transcription and

translation, also known as attenuation (Khan, n.d).

For example, bacteria found in our gut, ​Escherichia coli need amino acids to survive,

because proteins must be made. One of the amino acids that they need, is tryptophan. Once

tryptophan is available in the environment, E. coli takes it up and uses it to build proteins. In fact,

E. coli can also make its own tryptophan using enzymes that are encoded by five genes. These

five genes are located next to each other in what is called the trp operon (Khan, n.d). The trp

operon more specifically includes five genes that encodes the genes for the synthesis of

tryptophan along with RNA promoter site, and a binding site for a repressor protein (California

State University, n.d). Through the the process of transcription, the genes of the trp operon are

transcribed as a single mRNA (California State University, n.d). When the DNA binds to the

repressor, it keeps the operon from being transcribed. This is done when the repressor physically

gets in the way of the RNA polymerase, which is the transcribed enzyme (Khan, n.d). The gene

known as trpR is the gene that encodes the trp repressor protein, located n the bacterial
TRP OPERON MUTATIONS ASSIGNMENT 2

chromosome (California State University, n.d). When the DNA of the operator binds with the

repressor, it prevents the operon from being transcribed, by physically getting in the way of the

RNA polymerase, which is the transcription enzyme (Khan, n.d). However when tryptophan is

present the trp repressor does not bind with the DNA. Instead transcription is blocked. In

addition, when tryptophan is present, it attaches to the repressor molecules and makes their shape

more active. An example of a corepressor is when a small molecule like tryptophan switches a

repressor into its active state. However, when the trp repressor is inactive, it means that there are

little to no tryptophan left in the cell. As a result, trp operon is transcribed by RNA polymerase

because it does not attach to the DNA or block transcription.

On the contrary, the lac operon contains three genes known as the lacZ, lacY, and lacA.

Just as the trp operon, these genes are transcribed as a single mRNA, under the control of a

promoter (khan, n.d). In addition to the various genes, the lac operon contains many regulatory

DNA sequences. These regions of DNA are where regulatory proteins can bind, which controls

the transcription of an operon (Khan, n.d). The RNA polymerase which is an enzyme that

performs transcription, binds to the promoter site. Furthermore, the operator is a negative

regulatory site bound to the repressor protein (Farlex, n.d). The CAP binding site is a positive

regulatory site that is bound by the catabolite activator protein (Khan, n.d). In more details the

lac repressor, is a protein that is in charge of inhibiting transcription of the lac operon (Unknown

author, n.d). This is done when the operator binded, which overlaps the promoter (Unknown

author, n.d). Once bounded, the lac repressor gets in the way of RNA polymerases way,

utimarley keeping it from transcribing the operon (Unknown author, n.d).

TRP OPERON MUTATIONS ASSIGNMENT 3

When lactose is not available, the lac repressor binds to the operator, which prevents

transcription by RNA polymerase (Khan, n.d). However, when lactose is present, the lac

repressor cannot bind to the DNA. Instead it floats off the operator, which clears the way for

RNA polymerase to transcribe the operon (Khan, n.d). The change of the lac repressor is caused

by allolactose, an isomer of lactose (Khan, n.d). Allolactose binds to the lac repressor and makes

it change shape so it is longer possible to bind to the DNA. Allolactose is an inducer, which is a

small molecule that triggers expression of a gene or operon.

Based on your understanding of gene regulation in the cell and the function of the trp

operon, describe how this process would be affected if there was a mutation in the operator

region so that the operator could not carry out its function.

The trp operon includes five genes that encode enzymes needed for tryptophan

biosynthesis, along with a promoter and an operator. The genes of the trp operon are transcribed

as a single mRNA.​ This stretch of DNA is recognized by a regulatory protein known as the trp

repressor. When the repressor binds to the DNA of the operator, it keeps the operon from being

transcribed by physically getting in the way of RNA polymerase, the transcription enzyme.

However, if there was a mutation in the operator region so that the operator could not carry out

the function, then after when the trp repressor proteins are bounded to the tryptophan molecules,

they will lose the ability to bind to the operator site. Even through there is available repressor

protein, the trp operon will continue to go through transcription by transcribing genes into RNA

transcript. In conclusion the cell will be negatively impacted because energies and raw material

will be wasted.
TRP OPERON MUTATIONS ASSIGNMENT 4

What would be the result in this process of a genetic mutation that altered the shape of the

trp repressor protein?

The result of a genetic mutation that causes the shape of the trp repressor protein to change will

result in the repressor not being able to bind properly with tryptophan. As a result an excess

amount of tryptophan can be produced, due to the fact that the repressor is unable to bind with

the operator. As a result RNA polymerase will continuously bind to the promoter creating more

tryptophan. Furthermore, allosteric sites can also be changed too, resulting in a challenging

process in tryptophan molecules and corepressor combination. Overall a mutation would not be

essential because the trp operon will eventually lose ability to control the gene repression, thus

the cells will continue producing a lot of tryptophan molecules.

TRP OPERON MUTATIONS ASSIGNMENT 5

References

California State University. (n.d.). The trp Operon. Retrieved April 21, 2018, from

https://www.csun.edu/~hcbio027/biotechnology/lec4a/trp.html

Farlex. (n.d.). Operator site. Retrieved April 21, 2018, from

https://medical-dictionary.thefreedictionary.com/operator site

Khan, S. (n.d.). The trp operon. Retrieved April 21, 2018, from

https://www.khanacademy.org/science/biology/gene-regulation/gene-regulation-in-bacter

ia/

a/the-trp-operon

Khan, S. (n.d.). The lac operon. Retrieved April 21, 2018, from

https://www.khanacademy.org/science/biology/gene-regulation/gene-regulation-in-bacter

ia/a/the-lac-operon

The Operon. (2014, April 21). Retrieved from http://www.biology-pages.info/L/LacOperon.html