TRP OPERON

The trp operon encodes the genes for the synthesis of tryptophan. This
cluster of genes, like the lac operon, is regulated by a repressor that binds to
the operator sequences. The activity of the trp repressor for binding the
operator region is enhanced when it binds tryptophan; in this capacity,
tryptophan is known as a corepressor. Since the activity of the trp repressor
is enhanced in the presence of tryptophan, the rate of expression of the trp
operon is graded in response to the level of tryptophan in the cell.

Expression of the trp operon is also regulated by attenuation. The

attenuator region, which is composed of sequences found within the
transcribed RNA, is involved in controlling transcription from the operon
after RNA polymerase has initiated synthesis. The attenuator of sequences of
the RNA is found near the 5' end of the RNA termed the leader region of the
RNA. The leader sequences are located prior to the start of the coding region
for the first gene of the operon (the trpE gene). The attenuator region
contains codons for a small leader polypeptide that contains tandem
tryptophan codons. This region of the RNA is also capable of forming
several different stable stem-loop structures.

Depending on the level of tryptophan in the cell---and hence the level

of charged trp-tRNAs---the position of ribosome on the leader polypeptide
and the rate at which they are translating allows different stem-loops to
form. If tryptophan is abundant, the ribosome prevents stem-loop 1-2 from
forming and thereby favors stem-loop 3-4. The latter is found near a region
rich in uracil and acts as the transcriptional terminator loop as described in
the RNA synthesis page. Consequently, RNA polymerase is dislodged from
the template.

The operons coding for genes necessary for the synthesis of a number
of other amino acids are also regulated by this attenuation mechanism. It
should be clear, however, that this type of transcriptional regulation is not
feasible for eukaryotic cells.
The trp operon is an example of a biosynthetic operon whose expression is
regulated by an effector:

Trp Operon - and example of a repressible operon

• five genes (trpA, trpB, trpC, trpD, and trpE) involved in the
production of the amino acid tryptophan
• another gene (trpR) produces an inactive repressor protein
• accumulation of the end product (tryptophan) represses synthesis of
the enzymes
o tryptophan binds to the inactive repressor protein at an
allosteric site
o the conformation changes and the repressor + tryptophan
complex binds to the operator, repressing the operon
• tryptophan can accumulate due to internal production or from external
sources
o remember, E. coli is found in the intestines of humans so if you
eat a tryptophan-rich meal, this will accumulate in the bacteria
and turn off the operon
The operon consists of 5 structural genes that code for the three enzymes
required to convert chorismic acid into tryptophan:

• Anthranilate synthetase
component I - encoded by trpE
component II - encoded by trpD

• N-(5'-phosphoribosyl)-anthranilate isomerase/Indole-3-glycerol
phosphate synthase - encoded by trpC

• Tryptophan synthase
b-subunit - encoded by trpB
a-subunit - encoded by trpA

 The operon also contains a gene coding for a short oligopeptide (trpL)
which functions in attenuation (below).
 Expression is regulated by the Trp repressor protein which is encoded
by the trpR gene.
 Unlike the lac operon, trpR is not adjacent to the operon. The
structural genes are located at minute 28 on the E. coli map; trpR is
located at minute 100.
 The TrpR repressor actually regulates the expression of two other
operons in addition to the trp operon.
 TrpR protein is unable to bind to the operator by itself. As long as
there is insufficient tryptophan in the cell, it will remain so.
 However, once the level of tryptophan builds up, then the Trp
repressor will block further transcription of the operon and, as a result,
the synthesis of the three enzymes will decline.
Attenuation
A straightforward prediction of the operon model of gene expression applied
to the case of the trp operon is that expression of the operon in trpR mutants
should be insensitive to added tryptophan. If there is no repressor to affect
transcription, then since it is repressor that senses the presence of
tryptophan, there should be no effect.

Expression of trpEDCBA is reduced by the addition of trypophan in trpR

mutants.

Further research established that this second level of tryptophan control

involved two components:

• tRNA, specifically tryptophanyl-tRNATrp, i.e. tRNATrp charged with

tryptophan.
• the trpL gene

trpL codes for a short 14 aa oligopeptide. It contains two consecutive trp

codons and therefore serves as a measure of the tryptophan supply in the
cell. If the supply is good, then the tRNA will be charged and the leader
peptide will be translated without problem. If the supply is inadequate, then
the tRNA will not be charged, and translation will stall at the trp codons.

How does this affect transcription of the trp operon?

The explanation derives from the observation that the trpL mRNA region
can adopt a number of different conformations. It contains several self-
complementary regions which can form a variety of stem-loop structures as
shown:
In one conformation, shown at the left, a typical bacterial transcription
terminator can form as a result of pairing between regions 3 (green box) & 4
(yellow box). In the other conformation, at right, the terminator is precluded
from forming because region 3 is now paired with region 2 (purple box)
forming an alternative structure.

Now, what governs the formation of the terminator or the anti-terminator?

Recall that transcription and translation can occur simultaneously in

bacteria. This means that the ribosome will attach to mRNA and is able to
influence the formation of secondary structures by the mRNA.

In the case of the trpL mRNA, if there is a plentiful supply of tryptophan,

then the ribosome follows right behind RNA polymerase until it is halted by
a stop codon. This permits formation of the terminator stem-loop which will
cause RNA polymerase to dissociate.
Notice the UGA stop codon at the bottom of region 1 in the sequence
diagram above.

If the supply of tryptophan is low, then tRNATrp will not be charged with
tryptophan, and the ribosome will stall waiting for a suitable tRNA to be
brought to the A site:

Because the ribosome stalls, regions 2 (purple) can now base-pair with
region 3 (green) as soon as both are transcribed. Formation of the anti-
terminator prevents formation of the terminator.
Sources:
• www.mun.ca/biochem/courses/3107/Topics/Trp_operon.html
• en.wikipedia.org/wiki/Operon
• www.ndsu.nodak.edu/instruct/
• en.wikipedia.org/wiki/Trp_operon
• science.nhmccd.edu/biol/operon.html
• bcs.whfreeman.com/thelifewire/ content/chp13/1302002.html
• mmbr.asm.org/cgi/content/abstract/67/3/3034
• socrates.acadiau.ca/courses/ biol/Microbiology/regulation.htm
• www.ncbi.nlm.nih.gov/pubmed/18374625

I am very much thankful to all the sources I would be ever grateful