Skip to main content
NCBI home page
Journal of Bacteriology logoLink to Journal of Bacteriology
. 1983 Jul;155(1):15–21. doi: 10.1128/jb.155.1.15-21.1983

Temperature-sensitive autolysis-defective mutants of Escherichia coli.

R E Harkness, E E Ishiguro
PMCID: PMC217645  PMID: 6134714

Abstract

Two independently isolated temperature-sensitive autolysis-defective mutants of Escherichia coli LD5 (thi lysA dapD) were characterized. The mutants were isolated by screening the survivors of a three-step enrichment process involving sequential treatments with bactericidal concentrations of D-cycloserine, benzyl-penicillin, and D-cycloserine at 42 degrees C. Cultures of the mutants underwent autolysis during beta-lactam treatment, D-cycloserine treatment, or diaminopimelic acid deprivation at 30 degrees C. The same treatments at 42 degrees C inhibited growth but did not induce lysis of the mutants. The minimum inhibitory concentrations of selected beta-lactam antibiotics and D-cycloserine were identical for the parent and mutant strains at both 30 and 42 degrees C. Both mutants failed to form colonies at 42 degrees C, and both gave rise to spontaneous temperature-resistant revertants. The revertants exhibited the normal lytic response when treated with D-cycloserine and beta-lactams or when deprived of diaminopimelic acid at 42 degrees C. The basis for the autolysis-defective phenotype of these mutants could not be determined. However, a nonspecific in vitro assay for peptidoglycan hydrolase activity in cell-free extracts indicated that both mutants were deficient in a peptidoglycan hydrolase. Both mutations were localized to the 56- to 61-min region of the E. coli chromosome by F' complementation.

Full text

PDF
15

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Goodell E. W., Schwarz U. Enzymes synthesizing and hydrolyzing murein in Escherichia coli. Topographical distribution over the cell envelope. Eur J Biochem. 1977 Nov 15;81(1):205–210. doi: 10.1111/j.1432-1033.1977.tb11942.x. [DOI] [PubMed] [Google Scholar]
  2. Hakenbeck R., Goodell E. W., Schwarz U. Compartmentalization of murein hydrolases in the envelope of Escherichia coli. FEBS Lett. 1974 Apr 1;40(2):261–264. doi: 10.1016/0014-5793(74)80240-1. [DOI] [PubMed] [Google Scholar]
  3. Hansen M. T., Pato M. L., Molin S., Fill N. P., von Meyenburg K. Simple downshift and resulting lack of correlation between ppGpp pool size and ribonucleic acid accumulation. J Bacteriol. 1975 May;122(2):585–591. doi: 10.1128/jb.122.2.585-591.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Hartmann R., Bock-Hennig S. B., Schwarz U. Murein hydrolases in the envelope of Escherichia coli. Properties in situ and solubilization from the envelope. Eur J Biochem. 1974 Jan 3;41(1):203–208. doi: 10.1111/j.1432-1033.1974.tb03261.x. [DOI] [PubMed] [Google Scholar]
  5. Hartmann R., Höltje J. V., Schwarz U. Targets of penicillin action in Escherichia coli. Nature. 1972 Feb 25;235(5339):426–429. doi: 10.1038/235426a0. [DOI] [PubMed] [Google Scholar]
  6. Ishiguro E. E., Ramey W. D. Stringent control of peptidoglycan biosynthesis in Escherichia coli K-12. J Bacteriol. 1976 Sep;127(3):1119–1126. doi: 10.1128/jb.127.3.1119-1126.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Kitano K., Tomasz A. Escherichia coli mutants tolerant to beta-lactam antibiotics. J Bacteriol. 1979 Dec;140(3):955–963. doi: 10.1128/jb.140.3.955-963.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  9. Markwell M. A., Haas S. M., Bieber L. L., Tolbert N. E. A modification of the Lowry procedure to simplify protein determination in membrane and lipoprotein samples. Anal Biochem. 1978 Jun 15;87(1):206–210. doi: 10.1016/0003-2697(78)90586-9. [DOI] [PubMed] [Google Scholar]
  10. Noguchi H., Matsuhashi M., Mitsuhashi S. Comparative studies of penicillin-binding proteins in Pseudomonas aeruginosa and Escherichia coli. Eur J Biochem. 1979 Oct;100(1):41–49. doi: 10.1111/j.1432-1033.1979.tb02031.x. [DOI] [PubMed] [Google Scholar]
  11. PRIMOSIGH J., PELZER H., MAASS D., WEIDEL W. Chemical characterization of mucopeptides released from the E. coli B cell wall by enzymic action. Biochim Biophys Acta. 1961 Jan 1;46:68–80. doi: 10.1016/0006-3002(61)90647-3. [DOI] [PubMed] [Google Scholar]
  12. Tomasz A., Albino A., Zanati E. Multiple antibiotic resistance in a bacterium with suppressed autolytic system. Nature. 1970 Jul 11;227(5254):138–140. doi: 10.1038/227138a0. [DOI] [PubMed] [Google Scholar]
  13. Tomasz A. The mechanism of the irreversible antimicrobial effects of penicillins: how the beta-lactam antibiotics kill and lyse bacteria. Annu Rev Microbiol. 1979;33:113–137. doi: 10.1146/annurev.mi.33.100179.000553. [DOI] [PubMed] [Google Scholar]
  14. van Heijenoort J., Parquet C., Flouret B., van Heijenoort Y. Envelope-bound N-acetylmuramyl-L-alanine amidase of Escherichia coli K 12. Purification and properties of the enzyme. Eur J Biochem. 1975 Oct 15;58(2):611–619. doi: 10.1111/j.1432-1033.1975.tb02412.x. [DOI] [PubMed] [Google Scholar]

Articles from Journal of Bacteriology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES