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PMID: 12029049 Published · ppublish English Journal Article Research Support, U.S. Gov't, P.H.S.

Yersinia enterocolitica type III secretion: mutational analysis of the yopQ secretion signal.

Journal of bacteriology ·Vol. 184 ·No. 12 ·2002-06-00 ·Pages 3321-8

Ramamurthi KS, Schneewind O

Abstract

Pathogenic Yersinia spp. secrete Yop proteins via the type III pathway. yopQ codons 1 to 15 were identified as a signal necessary and sufficient for the secretion of a fused reporter protein. Frameshift mutations that alter codons 2 to 15 with little alteration of yopQ mRNA sequence do not abolish type III transport, suggesting a model in which yopQ mRNA may provide a signal for secretion (D. M. Anderson and O. Schneewind, Mol. Microbiol. 31:1139-1148, 2001). In a recent study, the yopE signal was truncated to codons 1 to 12. All frameshift mutations introduced within the first 12 codons of yopE abolished secretion. Also, multiple synonymous mutations that changed the mRNA sequence of yopE codons 1 to 12 without altering the amino acid sequence did not affect secretion. These results favor a model whereby an N-terminal signal peptide initiates YopE into the type III pathway (S. A. Lloyd et al., Mol. Microbiol. 39:520-531, 2001). It is reported here that codons 1 to 10 of yopQ act as a minimal secretion signal. Further truncation of yopQ, either at codon 10 or at codon 2, abolished secretion. Replacement of yopQ AUG with either of two other start codons, UUG or GUG, did not affect secretion. However, replacement of AUG with CUG or AAA and initiating translation at the fusion site with npt did not permit Npt secretion, suggesting that the translation of yopQ codons 1 to 15 is a prerequisite for secretion. Frameshift mutations of yopQ codons 1 to 10, 1 to 11, and 1 to 12 abolished secretion signaling, whereas frameshift mutations of yopQ codons 1 to 13, 1 to 14, and 1 to 15 did not. Codon changes at yopQ positions 2 and 10 affected secretion signaling when placed within the first 10 codons but had no effect when positioned in the larger fusion of yopQ codons 1 to 15. An mRNA mutant of yopQ codons 1 to 10, generated by a combination of nine synonymous mutations, was defective in secretion signaling, suggesting that the YopQ secretion signal is not proteinaceous. A model is discussed whereby the initiation of YopQ polypeptide into the type III pathway is controlled by properties of yopQ mRNA.

MeSH Terms
3' Untranslated Regions Bacterial Outer Membrane Proteins/genetics,metabolism Base Sequence Codon DNA Mutational Analysis Frameshift Mutation Gene Expression Regulation, Bacterial Molecular Sequence Data Protein Biosynthesis RNA, Messenger/genetics Signal Transduction Yersinia enterocolitica/genetics,metabolism
Chemicals
3' Untranslated Regions Bacterial Outer Membrane Proteins Codon RNA, Messenger YopQ protein, Yersinia enterocolitica
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Ramamurthi Kumaran S
Committee on Microbiology, University of Chicago, Chicago, Illinois 60637, USA.
Schneewind Olaf
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43 references, click to expand
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Article Info
Journal
Journal of bacteriology
Abbr.
J Bacteriol
ISSN
0021-9193
DOI
10.1128/JB.184.12.3321-3328.2002
Published
2002-06-00
Pages
3321-8
Language
English
Region
United States
NLM ID
2985120R
PMCID
PMC135085
Subset
IM
Grants
NIAID NIH HHS · R01 AI042797 · United States
NIAID NIH HHS · AI42797 · United States
External Links
PubMed source DOI: 10.1128/JB.184.12.3321-3328.2002 PMC Full Text View journal details
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