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PMID: 2152897 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Genetic analysis of the pAD1 hemolysin/bacteriocin determinant in Enterococcus faecalis: Tn917 insertional mutagenesis and cloning.

Journal of bacteriology ·Vol. 172 ·No. 1 ·1990-01-00 ·Pages 155-63

Ike Y, Clewell DB, Segarra RA, Gilmore MS

Abstract

Thirty-seven nonhemolytic/nonbacteriocinogenic mutations in Enterococcus (Streptococcus) faecalis plasmid pAD1 were generated by Tn917 insertion. All were found to belong to one of two complementation classes. Each class of mutants secreted either hemolysin/bacteriocin (Hly/Bac) component A or L into the culture medium. DNA encoding Hly/Bac was cloned in Escherichia coli in which both components of the hemolysin were expressed individually and collectively. The region encoding components A and L was further defined by deletion analysis and physically mapped. A total of approximately 8.4 kilobases of pAD1 DNA were observed to be required for hemolysin expression. Hly/Bac activity of the wild-type and the inactive L substance was observed to be heat stable. Active Hly/Bac resulting from incubating separately secreted components A and L was also found to be heat stable. The results indicate that component A activates component L and that activated component L possesses the Hly/Bac activity. Component A was also observed to be associated with host immunity to the Hly/Bac.

MeSH Terms
Bacteriocins Cloning, Molecular DNA Transposable Elements Enterococcus faecalis/genetics,immunology Genetic Complementation Test Hemolysin Proteins/genetics Hemolysis Hot Temperature Mutation
Chemicals
Bacteriocins DNA Transposable Elements Hemolysin Proteins
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Ike Y
Department of Biologic and Materials Sciences, School of Dentistry, University of Michigan, Ann Arbor 48109.
Clewell D B
Segarra R A
Gilmore M S
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Article Info
Journal
Journal of bacteriology
Abbr.
J Bacteriol
ISSN
0021-9193
DOI
10.1128/jb.172.1.155-163.1990
Published
1990-01-00
Pages
155-63
Language
English
Region
United States
NLM ID
2985120R
PMCID
PMC208413
Subset
IM
Grants
PHS HHS · AL 10318 · United States
NIDCR NIH HHS · DE 02731 · United States
NIGMS NIH HHS · GM 33956 · United States
External Links
PubMed source DOI: 10.1128/jb.172.1.155-163.1990 PMC Full Text View journal details
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