DOI: 10.1073/pnas.171299298. The mechanism of tryptophan induction of tryptophanase operon expression: Tryptophan inhibits release factor-mediated cleavage of TnaC-peptidyl-tRNA <sup>Pro</sup>

The mechanism of tryptophan induction of tryptophanase operon expression: Tryptophan inhibits release factor-mediated cleavage of TnaC-peptidyl-tRNA Pro

10.1073/pnas.171299298

Crossref journal-article

Proceedings of the National Academy of Sciences

Proceedings of the National Academy of Sciences (341)

Abstract

Expression of the tryptophanase ( tna ) operon of Escherichia coli is regulated by catabolite repression and tryptophan-induced transcription antitermination. In a previous study, we reproduced the regulatory features of this operon observed in vivo by using an in vitro S-30 system. We also found that, under inducing conditions, the leader peptidyl-tRNA (TnaC-peptidyl-tRNA Pro ) is not cleaved; it accumulates in the S-30 reaction mixture. In this paper, we examine the requirements for TnaC-peptidyl-tRNA Pro accumulation and cleavage, in vitro . We show that this peptidyl-tRNA remains bound to the translating ribosome. Removal of free tryptophan and addition of release factor 1 or 2 leads to hydrolysis of TnaC-peptidyl-tRNA Pro and release of TnaC from the ribosome-mRNA complex. Release factor-mediated cleavage is prevented by the addition of tryptophan. TnaC of the ribosome-bound TnaC-peptidyl-tRNA Pro was transferable to puromycin. This transfer was also blocked by tryptophan. Tests with various tryptophan analogs as substitutes for tryptophan revealed the existence of strict structural requirements for tryptophan action. Our findings demonstrate that the addition of tryptophan to ribosomes bearing nascent TnaC-peptidyl-tRNA Pro inhibits both TnaC peptidyl-tRNA Pro hydrolysis and TnaC peptidyl transfer. The associated translating ribosome therefore remains attached to the leader transcript where it blocks Rho factor binding and subsequent transcription termination.

Bibliography

Gong, F., Ito, K., Nakamura, Y., & Yanofsky, C. (2001). The mechanism of tryptophan induction of tryptophanase operon expression: Tryptophan inhibits release factor-mediated cleavage of TnaC-peptidyl-tRNA Pro. Proceedings of the National Academy of Sciences, 98(16), 8997â9001.

Authors 4

Feng Gong (first)
Koichi Ito (additional)
Yoshikazu Nakamura (additional)
Charles Yanofsky (additional)

References 38 Referenced 111

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Dates

Type	When
Created	23 years, 1 month ago (July 26, 2002, 10:34 a.m.)
Deposited	3 years, 4 months ago (April 13, 2022, 2:46 a.m.)
Indexed	1 week ago (Aug. 19, 2025, 5:58 a.m.)
Issued	24 years, 1 month ago (July 24, 2001)
Published	24 years, 1 month ago (July 24, 2001)
Published Online	24 years, 1 month ago (July 24, 2001)
Published Print	24 years ago (July 31, 2001)

Funders 0

None

BibTeX

@article{Gong_2001, title={The mechanism of tryptophan induction of tryptophanase operon expression: Tryptophan inhibits release factor-mediated cleavage of TnaC-peptidyl-tRNA Pro}, volume={98}, ISSN={1091-6490}, url={http://dx.doi.org/10.1073/pnas.171299298}, DOI={10.1073/pnas.171299298}, number={16}, journal={Proceedings of the National Academy of Sciences}, publisher={Proceedings of the National Academy of Sciences}, author={Gong, Feng and Ito, Koichi and Nakamura, Yoshikazu and Yanofsky, Charles}, year={2001}, month=jul, pages={8997–9001} }

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