PMID: 2501788Jul 1, 1989Paper

Intracellular methicillin selection of Listeria monocytogenes mutants unable to replicate in a macrophage cell line

Proceedings of the National Academy of Sciences of the United States of America
A CamilliD A Portnoy

Abstract

To dissect the determinants of Listeria monocytogenes that are required for pathogenicity, we designed an intracellular selection protocol based on penicillin selection to isolate mutants defective for intracellular growth. Eight independent mutants obtained by insertion of Tn916 were isolated that were resistant to methicillin treatment following internalization by the J774 macrophage-like cell line. Seven mutants were absolutely defective for intracellular growth, whereas one showed abortive intracellular growth. The majority of the mutants were nonhemolytic and lacked a secreted 58-kDa polypeptide thought to be the L. monocytogenes hemolysin, listeriolysin O. Southern blot analysis indicated that one mutant contained a Tn916 insertion in hlyA, the listeriolysin O structural gene, which resulted in a truncated listeriolysin O polypeptide, whereas another mutant contained an insertion immediately upstream of hlyA, which resulted in reduced expression of listeriolysin O. The other mutants contained Tn916 insertions in genes other than hlyA, although all but one were nonhemolytic. Revertants isolated by their ability to grow within tissue culture cells regained hemolytic activity. These data show that intracellular methicillin s...Continue Reading

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Citations

Feb 6, 2010·Proceedings of the National Academy of Sciences of the United States of America·Sophie HelaineDavid W Holden
Dec 15, 1991·Proceedings of the National Academy of Sciences of the United States of America·K Y Leung, B B Finlay
Feb 21, 2002·Infection and Immunity·Lynne M Shetron-RamaNancy E Freitag
Oct 24, 2012·Journal of Biomedicine & Biotechnology·R StachowiakJ Kawiak
Jan 1, 1992·Free Radical Research Communications·A Haas, W Goebel
Jul 13, 2002·FEMS Microbiology Letters·Jorge Castro-GarzaFrederick D Quinn
Aug 16, 2002·Molecular Microbiology·Laurel L Lenz, Daniel A Portnoy
Dec 1, 1990·Applied and Environmental Microbiology·A R DattaJ Russell
Jun 1, 1997·Microbiology and Molecular Biology Reviews : MMBR·B B Finlay, S Falkow
Oct 18, 2003·Science·Mary O'RiordanDaniel A Portnoy

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