Skip to main content
PMC home page
Journal of Bacteriology logoLink to Journal of Bacteriology
. 1995 Jul;177(13):3818–3823. doi: 10.1128/jb.177.13.3818-3823.1995

Characterization of the lactococcal abiD1 gene coding for phage abortive infection.

J Anba 1, E Bidnenko 1, A Hillier 1, D Ehrlich 1, M C Chopin 1
PMCID: PMC177101  PMID: 7601848

Abstract

Lactococcal phage abortive infection (AbiD1) determined by plasmid pIL105 is active on both prolate- and small-isometric-head phages of the C6A and 936 phage groups, respectively, which are considered two different species.The Abi phenotype was found to be encoded by a single gene, designated abiD1. The abiD1-encoded protein (351 amino acids) does not show homology with any known protein and has a deduced isoelectric point of 10. It also possesses two helix-turn-helix structures and an unusually high content of asparagine, isoleucine, and lysine. A consensual promoter with a TGy extension to the -10 box was mapped 76 bp upstream of the start codon. Transcription initiated at this strong promoter stops at a terminator located 48 bp downstream from the promoter. The termination process is very efficient, and transcripts corresponding to the abiD1 gene were not visible in our experimental conditions with or without phage infection. Expression of abiD1 under the control of a T7 promoter induced a lag phase in Lactococcus lactis cell growth, suggesting that overproduction of AbiD1 could be toxic for the cells. AbiD1 protein was visualized in Escherichia coli by using a tightly controlled expression system.

Full Text

The Full Text of this article is available as a PDF (361.2 KB).

Selected References

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

  1. Altschul S. F., Gish W., Miller W., Myers E. W., Lipman D. J. Basic local alignment search tool. J Mol Biol. 1990 Oct 5;215(3):403–410. doi: 10.1016/S0022-2836(05)80360-2. [DOI] [PubMed] [Google Scholar]
  2. Barberis-Maino L., Berger-Bächi B., Weber H., Beck W. D., Kayser F. H. IS431, a staphylococcal insertion sequence-like element related to IS26 from Proteus vulgaris. Gene. 1987;59(1):107–113. doi: 10.1016/0378-1119(87)90271-x. [DOI] [PubMed] [Google Scholar]
  3. Bidnenko E., Ehrlich D., Chopin M. C. Phage operon involved in sensitivity to the Lactococcus lactis abortive infection mechanism AbiD1. J Bacteriol. 1995 Jul;177(13):3824–3829. doi: 10.1128/jb.177.13.3824-3829.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Chopin A., Chopin M. C., Moillo-Batt A., Langella P. Two plasmid-determined restriction and modification systems in Streptococcus lactis. Plasmid. 1984 May;11(3):260–263. doi: 10.1016/0147-619x(84)90033-7. [DOI] [PubMed] [Google Scholar]
  5. Chopin A. Organization and regulation of genes for amino acid biosynthesis in lactic acid bacteria. FEMS Microbiol Rev. 1993 Sep;12(1-3):21–37. doi: 10.1111/j.1574-6976.1993.tb00011.x. [DOI] [PubMed] [Google Scholar]
  6. Chung C. T., Miller R. H. A rapid and convenient method for the preparation and storage of competent bacterial cells. Nucleic Acids Res. 1988 Apr 25;16(8):3580–3580. doi: 10.1093/nar/16.8.3580. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Cluzel P. J., Chopin A., Ehrlich S. D., Chopin M. C. Phage abortive infection mechanism from Lactococcus lactis subsp. lactis, expression of which is mediated by an Iso-ISS1 element. Appl Environ Microbiol. 1991 Dec;57(12):3547–3551. doi: 10.1128/aem.57.12.3547-3551.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Coffey A. G., Fitzgerald G. F., Daly C. Cloning and characterization of the determinant for abortive infection of bacteriophage from lactococcal plasmid pCI829. J Gen Microbiol. 1991 Jun;137(6):1355–1362. doi: 10.1099/00221287-137-6-1355. [DOI] [PubMed] [Google Scholar]
  9. Delecluse A., Bourgouin C., Klier A., Rapoport G. Nucleotide sequence and characterization of a new insertion element, IS240, from Bacillus thuringiensis israelensis. Plasmid. 1989 Jan;21(1):71–78. doi: 10.1016/0147-619x(89)90088-7. [DOI] [PubMed] [Google Scholar]
  10. Devereux J., Haeberli P., Smithies O. A comprehensive set of sequence analysis programs for the VAX. Nucleic Acids Res. 1984 Jan 11;12(1 Pt 1):387–395. doi: 10.1093/nar/12.1part1.387. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Durmaz E., Higgins D. L., Klaenhammer T. R. Molecular characterization of a second abortive phage resistance gene present in Lactococcus lactis subsp. lactis ME2. J Bacteriol. 1992 Nov;174(22):7463–7469. doi: 10.1128/jb.174.22.7463-7469.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Gasson M. J. Plasmid complements of Streptococcus lactis NCDO 712 and other lactic streptococci after protoplast-induced curing. J Bacteriol. 1983 Apr;154(1):1–9. doi: 10.1128/jb.154.1.1-9.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Gautier M., Chopin M. C. Plasmid-Determined Systems for Restriction and Modification Activity and Abortive Infection in Streptococcus cremoris. Appl Environ Microbiol. 1987 May;53(5):923–927. doi: 10.1128/aem.53.5.923-927.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Glatron M. F., Rapoport G. Biosynthesis of the parasporal inclusion of Bacillus thuringiensis: half-life of its corresponding messenger RNA. Biochimie. 1972;54(10):1291–1301. doi: 10.1016/s0300-9084(72)80070-1. [DOI] [PubMed] [Google Scholar]
  15. Haandrikman A. J., van Leeuwen C., Kok J., Vos P., de Vos W. M., Venema G. Insertion elements on lactococcal proteinase plasmids. Appl Environ Microbiol. 1990 Jun;56(6):1890–1896. doi: 10.1128/aem.56.6.1890-1896.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Hill C., Miller L. A., Klaenhammer T. R. Nucleotide sequence and distribution of the pTR2030 resistance determinant (hsp) which aborts bacteriophage infection in lactococci. Appl Environ Microbiol. 1990 Jul;56(7):2255–2258. doi: 10.1128/aem.56.7.2255-2258.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Holo H., Nes I. F. High-Frequency Transformation, by Electroporation, of Lactococcus lactis subsp. cremoris Grown with Glycine in Osmotically Stabilized Media. Appl Environ Microbiol. 1989 Dec;55(12):3119–3123. doi: 10.1128/aem.55.12.3119-3123.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Jarvis A. W., Fitzgerald G. F., Mata M., Mercenier A., Neve H., Powell I. B., Ronda C., Saxelin M., Teuber M. Species and type phages of lactococcal bacteriophages. Intervirology. 1991;32(1):2–9. doi: 10.1159/000150179. [DOI] [PubMed] [Google Scholar]
  19. Kumar A., Malloch R. A., Fujita N., Smillie D. A., Ishihama A., Hayward R. S. The minus 35-recognition region of Escherichia coli sigma 70 is inessential for initiation of transcription at an "extended minus 10" promoter. J Mol Biol. 1993 Jul 20;232(2):406–418. doi: 10.1006/jmbi.1993.1400. [DOI] [PubMed] [Google Scholar]
  20. Martin C., Timm J., Rauzier J., Gomez-Lus R., Davies J., Gicquel B. Transposition of an antibiotic resistance element in mycobacteria. Nature. 1990 Jun 21;345(6277):739–743. doi: 10.1038/345739a0. [DOI] [PubMed] [Google Scholar]
  21. McLandsborough L. A., Kolaetis K. M., Requena T., McKay L. L. Cloning and characterization of the abortive infection genetic determinant abiD isolated from pBF61 of Lactococcus lactis subsp. lactis KR5. Appl Environ Microbiol. 1995 May;61(5):2023–2026. doi: 10.1128/aem.61.5.2023-2026.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Mollet B., Iida S., Shepherd J., Arber W. Nucleotide sequence of IS26, a new prokaryotic mobile genetic element. Nucleic Acids Res. 1983 Sep 24;11(18):6319–6330. doi: 10.1093/nar/11.18.6319. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Nücken E. J., Henschke R. B., Schmidt F. R. Nucleotide-sequence of insertion element IS15 delta IV from plasmid pBP11. DNA Seq. 1990;1(1):85–88. doi: 10.3109/10425179009041351. [DOI] [PubMed] [Google Scholar]
  24. Pages J. M., Lazdunski C. Maturation of exported proteins in Escherichia coli. Requirement for energy, site and kinetics of processing. Eur J Biochem. 1982 Jun;124(3):561–566. [PubMed] [Google Scholar]
  25. Polzin K. M., Shimizu-Kadota M. Identification of a new insertion element, similar to gram-negative IS26, on the lactose plasmid of Streptococcus lactis ML3. J Bacteriol. 1987 Dec;169(12):5481–5488. doi: 10.1128/jb.169.12.5481-5488.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Romero D. A., Klaenhammer T. R. Characterization of insertion sequence IS946, an Iso-ISS1 element, isolated from the conjugative lactococcal plasmid pTR2030. J Bacteriol. 1990 Aug;172(8):4151–4160. doi: 10.1128/jb.172.8.4151-4160.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Rouch D. A., Skurray R. A. IS257 from Staphylococcus aureus: member of an insertion sequence superfamily prevalent among gram-positive and gram-negative bacteria. Gene. 1989;76(2):195–205. doi: 10.1016/0378-1119(89)90160-1. [DOI] [PubMed] [Google Scholar]
  28. Schmitt C. K., Molineux I. J. Expression of gene 1.2 and gene 10 of bacteriophage T7 is lethal to F plasmid-containing Escherichia coli. J Bacteriol. 1991 Feb;173(4):1536–1543. doi: 10.1128/jb.173.4.1536-1543.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Simon D., Chopin A. Construction of a vector plasmid family and its use for molecular cloning in Streptococcus lactis. Biochimie. 1988 Apr;70(4):559–566. doi: 10.1016/0300-9084(88)90093-4. [DOI] [PubMed] [Google Scholar]
  30. 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]
  31. Studier F. W., Rosenberg A. H., Dunn J. J., Dubendorff J. W. Use of T7 RNA polymerase to direct expression of cloned genes. Methods Enzymol. 1990;185:60–89. doi: 10.1016/0076-6879(90)85008-c. [DOI] [PubMed] [Google Scholar]
  32. 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]
  33. Terzaghi B. E., Sandine W. E. Improved medium for lactic streptococci and their bacteriophages. Appl Microbiol. 1975 Jun;29(6):807–813. doi: 10.1128/am.29.6.807-813.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Wells J. M., Wilson P. W., Norton P. M., Gasson M. J., Le Page R. W. Lactococcus lactis: high-level expression of tetanus toxin fragment C and protection against lethal challenge. Mol Microbiol. 1993 Jun;8(6):1155–1162. doi: 10.1111/j.1365-2958.1993.tb01660.x. [DOI] [PubMed] [Google Scholar]
  35. van Rooijen R. J., Gasson M. J., de Vos W. M. Characterization of the Lactococcus lactis lactose operon promoter: contribution of flanking sequences and LacR repressor to promoter activity. J Bacteriol. 1992 Apr;174(7):2273–2280. doi: 10.1128/jb.174.7.2273-2280.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. van Rooijen R. J., de Vos W. M. Molecular cloning, transcriptional analysis, and nucleotide sequence of lacR, a gene encoding the repressor of the lactose phosphotransferase system of Lactococcus lactis. J Biol Chem. 1990 Oct 25;265(30):18499–18503. [PubMed] [Google Scholar]
  37. van de Guchte M., Kok J., Venema G. Gene expression in Lactococcus lactis. FEMS Microbiol Rev. 1992 Feb;8(2):73–92. doi: 10.1111/j.1574-6968.1992.tb04958.x. [DOI] [PubMed] [Google Scholar]
  38. van der Vossen J. M., van der Lelie D., Venema G. Isolation and characterization of Streptococcus cremoris Wg2-specific promoters. Appl Environ Microbiol. 1987 Oct;53(10):2452–2457. doi: 10.1128/aem.53.10.2452-2457.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]

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

RESOURCES