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PMID: 9324268 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Constitutive expression of fibronectin binding in Streptococcus pyogenes as a result of anaerobic activation of rofA.

Journal of bacteriology ·Vol. 179 ·No. 19 ·1997-10-00 ·Pages 6172-80

Fogg GC, Caparon MG

Abstract

Protein F is a fibronectin-binding surface protein of Streptococcus pyogenes (group A streptococcus) that mediates adherence to host cells. A gene product encoded by rofA activates transcription of the gene that encodes protein F (prtF) and was identified in a strain of S. pyogenes that expressed high levels of protein F under all conditions tested. Insertional inactivation of rofA in this strain results in a phenotype similar to that of other strains where high-level transcription of prtF occurs only in response to increased oxygen tension. In this study, we have compared the regulation of prtF and rofA in O2-regulated and constitutive strains in order to gain further insight into the function of rofA. Comparison of the prtF and rofA transcripts by S1 nuclease and primer extension assays indicated that the same promoters for each transcript are used in both O2-regulated and constitutive strains. However, analyses of rofA-lacZ reporter alleles revealed that a key difference between strains involves regulation of rofA itself. In O2-regulated strains, expression of rofA was elevated following culture under conditions of reduced O2 tension. However, a much more robust activation of rofA expression was observed when constitutive strains were grown under similar conditions. Exchange of reporter and rofA alleles between strains demonstrated that host genetic background, and not the sequence of the respective rofA allele or regulatory region, dictates the expression phenotype. Activation of rofA required RofA, and RofA was shown to bind specifically to DNA containing the promoters for rofA and prtF. Finally, overexpression of either allele of rofA caused constitutive expression of prtF regardless of host background. These data suggest a model where anaerobic expression of prtF in constitutive hosts is controlled at the level of transcription of rofA and implicate additional factors in this regulatory pathway.

MeSH Terms
Adhesins, Bacterial/genetics Amino Acid Sequence Anaerobiosis Bacterial Proteins Base Sequence DNA, Bacterial/genetics,metabolism Fibronectins/metabolism Gene Expression Regulation, Bacterial Genetic Vectors Molecular Sequence Data Oxygen/physiology Promoter Regions, Genetic RNA, Bacterial/genetics,metabolism RNA, Messenger/genetics,metabolism Streptococcus pyogenes/genetics,metabolism Trans-Activators/genetics,metabolism Transcription, Genetic
Chemicals
Adhesins, Bacterial Bacterial Proteins DNA, Bacterial Fibronectins RNA, Bacterial RNA, Messenger Trans-Activators fibronectin-binding proteins, bacterial rofA protein, Streptococcus Oxygen
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Fogg G C
Department of Molecular Microbiology, Washington University School of Medicine, St. Louis, Missouri 63110-1093, USA.
Caparon M G
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Article Info
Journal
Journal of bacteriology
Abbr.
J Bacteriol
ISSN
0021-9193
DOI
10.1128/jb.179.19.6172-6180.1997
Published
1997-10-00
Pages
6172-80
Language
English
Region
United States
NLM ID
2985120R
PMCID
PMC179524
Subset
IM
Grants
NIAID NIH HHS · AI38273 · United States
Databases
GENBANK
U53825
External Links
PubMed source DOI: 10.1128/jb.179.19.6172-6180.1997 PMC Full Text View journal details
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