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PMID: 9045840 Published · ppublish English Journal Article Research Support, U.S. Gov't, P.H.S.

Regulation of the Escherichia coli tna operon: nascent leader peptide control at the tnaC stop codon.

Journal of bacteriology ·Vol. 179 ·No. 5 ·1997-03-00 ·Pages 1774-9

Konan KV, Yanofsky C

Abstract

Expression of the tryptophanase (tna) operon of Escherichia coli is regulated by catabolite repression and by tryptophan-induced transcription antitermination at Rho-dependent termination sites in the leader region of the operon. Tryptophan induction is dependent on translation of a short leader peptide coding region, tnaC, that contains a single, crucial tryptophan codon. Recent studies suggest that during induction, the TnaC leader peptide acts in cis on the translating ribosome to inhibit its release at the tnaC stop codon. In the present study we use a tnaC-UGA-'lacZ construct lacking the tnaC-tnaA spacer region to analyze the effect of TnaC synthesis on the behavior of the ribosome that translates tnaC. The tnaC-UGA-'lacZ construct is not expressed significantly in the presence or absence of inducer. However, it is expressed in the presence of UGA suppressors, or when the structural gene for polypeptide release factor 3 is disrupted, or when wild-type tRNATrP is overproduced. In each situation, tnaC-UGA-'lacZ expression is reduced appreciably by the presence of inducing levels of tryptophan. Replacing the tnaC UGA stop codon with a sense codon allows considerable expression, which is also reduced, although to a lesser extent, by the addition of tryptophan. Inhibition by tryptophan is not observed when Trp codon 12 of tnaC is changed to a Leu codon. Overexpression of tnaC in trans from a multicopy plasmid prevents inhibition of expression by tryptophan. These results support the hypothesis that the TnaC leader peptide acts in cis to alter the behavior of the translating ribosome.

MeSH Terms
Amino Acid Sequence Bacterial Proteins/chemistry,genetics,physiology Base Sequence Codon, Terminator Culture Media Escherichia coli/enzymology,genetics Escherichia coli Proteins Gene Expression Regulation, Bacterial Molecular Sequence Data Mutagenesis, Site-Directed Operon Peptide Termination Factors/genetics,physiology Protein Sorting Signals/physiology Recombinant Proteins Rho Factor/genetics,physiology Suppression, Genetic Tryptophan/metabolism Tryptophanase/genetics
Chemicals
Bacterial Proteins Codon, Terminator Culture Media Escherichia coli Proteins Peptide Termination Factors Protein Sorting Signals Recombinant Proteins Rho Factor peptide-chain-release factor 3 tnaC protein, E coli Tryptophan Tryptophanase
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Konan K V
Department of Biological Sciences, Stanford University, California 94305-5020, USA.
Yanofsky C
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Article Info
Journal
Journal of bacteriology
Abbr.
J Bacteriol
ISSN
0021-9193
DOI
10.1128/jb.179.5.1774-1779.1997
Published
1997-03-00
Pages
1774-9
Language
English
Region
United States
NLM ID
2985120R
PMCID
PMC178893
Subset
IM
Grants
NIGMS NIH HHS · GM09738 · United States
External Links
PubMed source DOI: 10.1128/jb.179.5.1774-1779.1997 PMC Full Text View journal details
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