Abstract
A Lyt- mutant with reduced autolytic activity was isolated after Tn551 mutagenesis of the methicillin-susceptible Staphylococcus aureus laboratory strain RN450. The Lyt- phenotype could be transferred back into the parent and into a variety of other S. aureus strains by transduction of the transposon marker. Southern analysis has located the Tn551 insert to a 3.2-kb HindIII DNA fragment on the SmaI B fragment of the staphylococcal chromosome. The Lyt- phenotype included reduced rates of cell wall turnover and autolysis induced by detergent or methicillin treatment; however, the rate of methicillin-induced killing was not affected. Peptidoglycans prepared from the parental and mutant cells showed identical muropeptide compositions, as resolved by a high-resolution high-pressure liquid chromatography technique. On the other hand, LiCl extracts of the mutant cells contained reduced amounts of total protein and lower specific cell wall-degrading activity compared with those of extracts of parental cells. The profile of bacteriolytic enzymes as detected by sodium dodecyl sulfate-polyacrylamide gel electrophoresis showed multiple band differences between mutant and parental cells; a major lytic band with properties characteristic of the staphylococcal endo-beta-N-acetylglucosaminidase was completely absent from the Lyt- cells. The Lyt- phenotype transduced into a series of methicillin-resistant strains of both homogeneous and heterogeneous phenotypes caused only a modest decrease in the level of methicillin resistance, as determined by population analysis.
Full text
PDF
Images in this article
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Bradford M. M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976 May 7;72:248–254. doi: 10.1006/abio.1976.9999. [DOI] [PubMed] [Google Scholar]
- Chatterjee A. N., Wong W., Young F. E., Gilpin R. W. Isolation and characterization of a mutant of Staphylococcus aureus deficient in autolytic activity. J Bacteriol. 1976 Mar;125(3):961–967. doi: 10.1128/jb.125.3.961-967.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Huff E., Silverman C. S., Adams N. J., Awkard W. S. Extracellular cell wall lytic enzyme from Staphylococcus aureus: purification and partial characterization. J Bacteriol. 1970 Sep;103(3):761–769. doi: 10.1128/jb.103.3.761-769.1970. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Jayaswal R. K., Lee Y. I., Wilkinson B. J. Cloning and expression of a Staphylococcus aureus gene encoding a peptidoglycan hydrolase activity. J Bacteriol. 1990 Oct;172(10):5783–5788. doi: 10.1128/jb.172.10.5783-5788.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Koyama T., Yamada M., Matsuhashi M. Formation of regular packets of Staphylococcus aureus cells. J Bacteriol. 1977 Mar;129(3):1518–1523. doi: 10.1128/jb.129.3.1518-1523.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Leclerc D., Asselin A. Detection of bacterial cell wall hydrolases after denaturing polyacrylamide gel electrophoresis. Can J Microbiol. 1989 Aug;35(8):749–753. doi: 10.1139/m89-125. [DOI] [PubMed] [Google Scholar]
- Maidhof H., Reinicke B., Blümel P., Berger-Bächi B., Labischinski H. femA, which encodes a factor essential for expression of methicillin resistance, affects glycine content of peptidoglycan in methicillin-resistant and methicillin-susceptible Staphylococcus aureus strains. J Bacteriol. 1991 Jun;173(11):3507–3513. doi: 10.1128/jb.173.11.3507-3513.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Murakami K., Tomasz A. Involvement of multiple genetic determinants in high-level methicillin resistance in Staphylococcus aureus. J Bacteriol. 1989 Feb;171(2):874–879. doi: 10.1128/jb.171.2.874-879.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
- PARK J. T., JOHNSON M. J. A submicrodetermination of glucose. J Biol Chem. 1949 Nov;181(1):149–151. [PubMed] [Google Scholar]
- Singer H. J., Wise E. M., Jr, Park J. T. Properties and purification of N-acetylmuramyl-L-alanine amidase from Staphylococcus aureus H. J Bacteriol. 1972 Nov;112(2):932–939. doi: 10.1128/jb.112.2.932-939.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Sugai M., Akiyama T., Komatsuzawa H., Miyake Y., Suginaka H. Characterization of sodium dodecyl sulfate-stable Staphylococcus aureus bacteriolytic enzymes by polyacrylamide gel electrophoresis. J Bacteriol. 1990 Nov;172(11):6494–6498. doi: 10.1128/jb.172.11.6494-6498.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Sugai M., Koike H., Hong Y. M., Miyake Y., Nogami R., Suginaka H. Purification of a 51 kDa endo-beta-N-acetylglucosaminidase from Staphylococcus aureus. FEMS Microbiol Lett. 1989 Oct 15;52(3):267–272. doi: 10.1016/0378-1097(89)90209-7. [DOI] [PubMed] [Google Scholar]
- Tipper D. J. Mechanism of autolysis of isolated cell walls of Staphylococcus aureus. J Bacteriol. 1969 Feb;97(2):837–847. doi: 10.1128/jb.97.2.837-847.1969. [DOI] [PMC free article] [PubMed] [Google Scholar]
- de Jonge B. L., de Lencastre H., Tomasz A. Suppression of autolysis and cell wall turnover in heterogeneous Tn551 mutants of a methicillin-resistant Staphylococcus aureus strain. J Bacteriol. 1991 Feb;173(3):1105–1110. doi: 10.1128/jb.173.3.1105-1110.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]