Prokaryotic Gene Regulation: Discussion #9 worksheet

1. What is gene regulation? What is the goal of gene regulation?

Gene regulation is controlling when and where genes are expressed. It controls how much of a functional protein is
present in the cell. The goal is to adjust the production levels of proteins to conserve a cells resources if a
substance is already available in the surroundings.
2. What is an operon? What is the goal of an operon (why have them)? What is the product of transcription of an
operon?
An operon is combined of the operator, the promoter and the genes they control. The goal of an operon is to adjust
the production levels of certain enzymes. If an operon undergoes transcription it will produce a particular enzyme
depending on the DNA stretch it contains.
3. A typical operon will have at least the following components/DNA sequences. For each component, specify the
general role of each in the operon and which if any proteins will bind to it.
promote
r

operator

Gene 1

Gene 2

Gene 3

Gene 4

Terminator

Promoter: The promoter is the DNA sequence where RNA polymerase can attach to initiate transcription
Operator: The operator is the switch for a segment of DNA. It controls the access of RNA polymerase to the genes.
A repressor can bind to the operator to block the attachment of RNA polymerase to turn it off.
Genes: The genes are stretches of DNA that contain instructions for building certain enzymes or proteins,
Terminator: The terminator has the purpose of signaling the end of transcription
4. The trp operon contains 5 genes (trp a, trp b, trp c, trp d and trp e), which code for the enzymes in the metabolic
pathway necessary to make tryptophan in the E. coli cell. The goal of the trp operon is to make tryptophan =
when the trp operon is on the cell will make the enzymes to make tryptophan and will thus make tryptophan. RNA
Pol, the trp repressor protein and tryptophan control expression of the trp operon.

promote
r

operator

trpE

trpD

trpC

trpB

trpA

Terminator

a. When tryptophan is available from the environment, would you expect the trp operon to be on or off?
_____Off_________
b. Explain your answer to (a) by describing where/how you would expect to find RNA Pol and the trp repressor
protein in this scenario (trp + environment). The trp repressor protein is active and binds to the operator which
blocks transcription because RNA polymerase cannot attach.
c. When tryptophan is not available from the environment, would you expect the trp operon to be on or off?
______On_____________
d. Explain your answer to (c) by describing where/how you would expect to find RNA Pol and the trp repressor
protein in this scenario (trp environment). The repressor is inactive and not bound to the operator. RNA
polymerase attaches to the promoter and begins transcription.
e. Is the trp operon inducible (turned on when needed) or repressible (turned off when not needed?)
______Repressible____________
f. Is the trp operon an example of positive or negative transcriptional regulation? ___________Negative_____
5. The lac operon contains 3 genes (lacZ, lacY and lacA), which code for the enzymes involved in the uptake and
breakdown of the disaccharide lactose into two monosaccharides. The goal of the lac operon is to breakdown
lactose into galactose and glucose (so they can be used as an energy source for glycolysis) = when the lac operon

is on the cell will make the enzymes to breakdown lactose and thus breakdown lactose. RNA Pol, the lac repressor
protein, CAP, lactose, glucose and cAMP control expression of the lac operon.

promote
r

CPS

operator

lacZ

lacY

lacA

Terminator

a. Would you expect the lac operon to be on or off, high or low when there is glucose available, but no lactose?
______Off_________
b. Explain your answer to (a) by describing where/how you would expect to find RNA Pol, the lac repressor protein
and the CAP protein in this scenario (glucose +, lactose - environment). Lac repressor is active and bound to the
operater. RNA pol cannot attach
c. Would you expect the lac operon to be on or off, high or low when there is lactose available, but no glucose?
________High____________
d. Explain your answer to (a) by describing where/how you would expect to find RNA Pol, the lac repressor protein
and the CAP protein in this scenario (glucose -, lactose + environment). High level of cAMP will activate CAP which
will bind to the DNA upstream of the promoter. This increases affinity of RNA pol to bind which will stimulate gene
expression. Lac repressor is inactive.
e. Would you expect the lac operon to be on or off, high or low when there is both lactose and glucose available?
_______Low__________
f. Explain your answer to (a) by describing where/how you would expect to find RNA Pol, the lac repressor protein
and the CAP protein in this scenario (glucose +, lactose + environment). cAMP is scarce so the CAP protein is
inactive and detaches from the operon. Because of this, RNA pol binds less efficiently so that transcription occurs
only at a low level. Lac repressor protein is also inactive.
g. Would you expect the lac operon to be on or off, high or low when there is neither lactose or glucose
available? ___Off________
h. Explain your answer to (a) by describing where/how you would expect to find RNA Pol, the lac repressor protein
and the CAP protein in this scenario (glucose -, lactose - environment). The repressor binds to the operator which
makes RNA pol unable to bind and unable to transcribe the gene.
e. Is the lac operon inducible (turned on when needed) or repressible (turned off when not needed?)
___Inducible______
f. Is the lac operon an example of positive or negative transcriptional regulation? _Both- negative because
allolactose derepresses the operon and positive because when CAP binds it switches transcription on _____