Skip to main content
PMC home page
Journal of Bacteriology logoLink to Journal of Bacteriology
. 1993 Feb;175(4):1038–1042. doi: 10.1128/jb.175.4.1038-1042.1993

The missing link in phage lysis of gram-positive bacteria: gene 14 of Bacillus subtilis phage phi 29 encodes the functional homolog of lambda S protein.

M Steiner 1, W Lubitz 1, U Bläsi 1
PMCID: PMC193017  PMID: 8432697

Abstract

In most bacteriophages of gram-negative bacteria, the phage endolysin is released to its murein substrate through a lesion in the inner membrane. The lesion is brought about by a second phage-encoded lysis function. For the first time, we present evidence that the same strategy is elaborated by a phage of a gram-positive bacterium. Thus, there appears to be an evolutionarily conserved lysis pathway for most phages whether their host bacterium is gram negative or gram positive. Phage phi 29 gene 14, the product of which is required for efficient lysis of Bacillus subtilis, was cloned in Escherichia coli. Production of protein 14 in E. coli resulted in cell death, whereas production of protein 14 concomitantly with the phi 29 lysozyme or unrelated murein-degrading enzymes led to lysis, suggesting that membrane-bound protein 14 induces a nonspecific lesion in the cytoplasmic membrane.

Full text

PDF

Images in this article

1041Image
on p.1041

Selected References

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

  1. Bienkowska-Szewczyk K., Lipinska B., Taylor A. The R gene product of bacteriophage lambda is the murein transglycosylase. Mol Gen Genet. 1981;184(1):111–114. doi: 10.1007/BF00271205. [DOI] [PubMed] [Google Scholar]
  2. Bläsi U., Chang C. Y., Zagotta M. T., Nam K. B., Young R. The lethal lambda S gene encodes its own inhibitor. EMBO J. 1990 Apr;9(4):981–989. doi: 10.1002/j.1460-2075.1990.tb08200.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bläsi U., Nam K., Hartz D., Gold L., Young R. Dual translational initiation sites control function of the lambda S gene. EMBO J. 1989 Nov;8(11):3501–3510. doi: 10.1002/j.1460-2075.1989.tb08515.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Bläsi U., Nam K., Lubitz W., Young R. Translational efficiency of phi X174 lysis gene E is unaffected by upstream translation of the overlapping gene D reading frame. J Bacteriol. 1990 Oct;172(10):5617–5623. doi: 10.1128/jb.172.10.5617-5623.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Bonovich M. T., Young R. Dual start motif in two lambdoid S genes unrelated to lambda S. J Bacteriol. 1991 May;173(9):2897–2905. doi: 10.1128/jb.173.9.2897-2905.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Carrascosa J. L., Camacho A., Moreno F., Jiménez F., Mellado R. P., Viñuela E., Salas M. Bacillus subtilis phage phi29. Characterization of gene products and functions. Eur J Biochem. 1976 Jul 1;66(2):229–241. doi: 10.1111/j.1432-1033.1976.tb10512.x. [DOI] [PubMed] [Google Scholar]
  7. Garrett J. M., Young R. Lethal action of bacteriophage lambda S gene. J Virol. 1982 Dec;44(3):886–892. doi: 10.1128/jvi.44.3.886-892.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Garrett J., Bruno C., Young R. Lysis protein S of phage lambda functions in Saccharomyces cerevisiae. J Bacteriol. 1990 Dec;172(12):7275–7277. doi: 10.1128/jb.172.12.7275-7277.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Garrett J., Fusselman R., Hise J., Chiou L., Smith-Grillo D., Schulz J., Young R. Cell lysis by induction of cloned lambda lysis genes. Mol Gen Genet. 1981;182(2):326–331. doi: 10.1007/BF00269678. [DOI] [PubMed] [Google Scholar]
  10. Garvey K. J., Saedi M. S., Ito J. Nucleotide sequence of Bacillus phage phi 29 genes 14 and 15: homology of gene 15 with other phage lysozymes. Nucleic Acids Res. 1986 Dec 22;14(24):10001–10008. doi: 10.1093/nar/14.24.10001. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Inouye M., Arnheim N., Sternglanz R. Bacteriophage T7 lysozyme is an N-acetylmuramyl-L-alanine amidase. J Biol Chem. 1973 Oct 25;248(20):7247–7252. [PubMed] [Google Scholar]
  12. Ito J. Bacteriophage phi29 terminal protein: its association with the 5' termini of the phi29 genome. J Virol. 1978 Dec;28(3):895–904. doi: 10.1128/jvi.28.3.895-904.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Jobling M. G., Holmes R. K. Construction of vectors with the p15a replicon, kanamycin resistance, inducible lacZ alpha and pUC18 or pUC19 multiple cloning sites. Nucleic Acids Res. 1990 Sep 11;18(17):5315–5316. doi: 10.1093/nar/18.17.5315. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Mindich L., Lehman J. Cell wall lysin as a component of the bacteriophage phi 6 virion. J Virol. 1979 May;30(2):489–496. doi: 10.1128/jvi.30.2.489-496.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Montag D., Degen M., Henning U. Nucleotide sequence of gene t (lysis gene) of the E. coli phage T4. Nucleic Acids Res. 1987 Aug 25;15(16):6736–6736. doi: 10.1093/nar/15.16.6736. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Nam K., Bläsi U., Zagotta M. T., Young R. Conservation of a dual-start motif in P22 lysis gene regulation. J Bacteriol. 1990 Jan;172(1):204–211. doi: 10.1128/jb.172.1.204-211.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Paces V., Vlcek C., Urbánek P., Hostomský Z. Nucleotide sequence of the right early region of Bacillus subtilis phage PZA completes the 19366-bp sequence of PZA genome. Comparison with the homologous sequence of phage phi 29. Gene. 1986;44(1):115–120. doi: 10.1016/0378-1119(86)90049-1. [DOI] [PubMed] [Google Scholar]
  18. Raab R., Neal G., Garrett J., Grimaila R., Fusselman R., Young R. Mutational analysis of bacteriophage lambda lysis gene S. J Bacteriol. 1986 Sep;167(3):1035–1042. doi: 10.1128/jb.167.3.1035-1042.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Raab R., Neal G., Sohaskey C., Smith J., Young R. Dominance in lambda S mutations and evidence for translational control. J Mol Biol. 1988 Jan 5;199(1):95–105. doi: 10.1016/0022-2836(88)90381-6. [DOI] [PubMed] [Google Scholar]
  20. Rennell D., Poteete A. R. Phage P22 lysis genes: nucleotide sequences and functional relationships with T4 and lambda genes. Virology. 1985 May;143(1):280–289. doi: 10.1016/0042-6822(85)90115-1. [DOI] [PubMed] [Google Scholar]
  21. Riede I. Lysis gene t of T-even bacteriophages: evidence that colicins and bacteriophage genes have common ancestors. J Bacteriol. 1987 Jul;169(7):2956–2961. doi: 10.1128/jb.169.7.2956-2961.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Saedi M. S., Garvey K. J., Ito J. Cloning and purification of a unique lysozyme produced by Bacillus phage phi 29. Proc Natl Acad Sci U S A. 1987 Feb;84(4):955–958. doi: 10.1073/pnas.84.4.955. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Shearman C., Underwood H., Jury K., Gasson M. Cloning and DNA sequence analysis of a Lactococcus bacteriophage lysin gene. Mol Gen Genet. 1989 Aug;218(2):214–221. doi: 10.1007/BF00331271. [DOI] [PubMed] [Google Scholar]
  24. Studier F. W., Moffatt B. A. Use of bacteriophage T7 RNA polymerase to direct selective high-level expression of cloned genes. J Mol Biol. 1986 May 5;189(1):113–130. doi: 10.1016/0022-2836(86)90385-2. [DOI] [PubMed] [Google Scholar]
  25. Studier F. W. Use of bacteriophage T7 lysozyme to improve an inducible T7 expression system. J Mol Biol. 1991 May 5;219(1):37–44. doi: 10.1016/0022-2836(91)90855-z. [DOI] [PubMed] [Google Scholar]
  26. Tsugita A., Inouye M. Complete primary structure of phage lysozyme from Escherichia coli T4. J Mol Biol. 1968 Oct 14;37(1):201–212. doi: 10.1016/0022-2836(68)90083-1. [DOI] [PubMed] [Google Scholar]
  27. Vlcek C., Paces V. Nucleotide sequence of the late region of Bacillus phage phi 29 completes the 19,285-bp sequence of phi 29 genome. Comparison with the homologous sequence of phage PZA. Gene. 1986;46(2-3):215–225. doi: 10.1016/0378-1119(86)90406-3. [DOI] [PubMed] [Google Scholar]
  28. Wilson D. B., Okabe A. A second function of the S gene of bacteriophage lambda. J Bacteriol. 1982 Dec;152(3):1091–1095. doi: 10.1128/jb.152.3.1091-1095.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Young R. Bacteriophage lysis: mechanism and regulation. Microbiol Rev. 1992 Sep;56(3):430–481. doi: 10.1128/mr.56.3.430-481.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Zagotta M. T., Wilson D. B. Oligomerization of the bacteriophage lambda S protein in the inner membrane of Escherichia coli. J Bacteriol. 1990 Feb;172(2):912–921. doi: 10.1128/jb.172.2.912-921.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]

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

RESOURCES