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. 1988 Sep;170(9):4343–4352. doi: 10.1128/jb.170.9.4343-4352.1988

Regulation of the pAD1 sex pheromone response in Enterococcus faecalis: construction and characterization of lacZ transcriptional fusions in a key control region of the plasmid.

K E Weaver 1, D B Clewell 1
PMCID: PMC211448  PMID: 2842316

Abstract

Strains carrying the Enterococcus (formerly Streptococcus) faecalis plasmid pAD1 responded to exogenous sex pheromone by inducing a number of gene products which facilitated mating. A 7-kilobase region of pAD1 was identified which contained genes that are important for the regulation of this response. Using the transposon Tn917-lac delivery vector pTV32Ts, we generated a number of fusions that allowed us to examine transcription in this region. At least three transcriptional units were identified by grouping fusions by their phenotype, direction of transcription, and response to pheromone. Transcription from one set of fusions was sensitive to the presence of pheromone. Analysis of the patterns of protein production previously shown to be induced in the presence of pheromone provided more information on the function of the genes of interest. We postulate the existence of two negative regulatory proteins that act coordinately to repress the pheromone response, one of which may be involved in sensing or transmitting the pheromone signal, and at least one positive regulatory protein whose synthesis is dependent on the presence of pheromone. In addition, the isolation of a relatively small deletion mutant capable of producing cAD1. the pheromone specific for pAD1-containing cells, indicates that a factor(s) that is important for the shutdown of endogenous pheromone is also present in this region.

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Selected References

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  1. Clewell D. B., An F. Y., White B. A., Gawron-Burke C. Streptococcus faecalis sex pheromone (cAM373) also produced by Staphylococcus aureus and identification of a conjugative transposon (Tn918). J Bacteriol. 1985 Jun;162(3):1212–1220. doi: 10.1128/jb.162.3.1212-1220.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Clewell D. B., Brown B. L. Sex pheromone cAD1 in Streptococcus faecalis: induction of a function related to plasmid transfer. J Bacteriol. 1980 Aug;143(2):1063–1065. doi: 10.1128/jb.143.2.1063-1065.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Clewell D. B., Tomich P. K., Gawron-Burke M. C., Franke A. E., Yagi Y., An F. Y. Mapping of Streptococcus faecalis plasmids pAD1 and pAD2 and studies relating to transposition of Tn917. J Bacteriol. 1982 Dec;152(3):1220–1230. doi: 10.1128/jb.152.3.1220-1230.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Clewell D. B., Yagi Y., Dunny G. M., Schultz S. K. Characterization of three plasmid deoxyribonucleic acid molecules in a strain of Streptococcus faecalis: identification of a plasmid determining erythromycin resistance. J Bacteriol. 1974 Jan;117(1):283–289. doi: 10.1128/jb.117.1.283-289.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Dempsey W. B. Transcript analysis of the plasmid R100 traJ and finP genes. Mol Gen Genet. 1987 Oct;209(3):533–544. doi: 10.1007/BF00331160. [DOI] [PubMed] [Google Scholar]
  6. Dunny G. M., Brown B. L., Clewell D. B. Induced cell aggregation and mating in Streptococcus faecalis: evidence for a bacterial sex pheromone. Proc Natl Acad Sci U S A. 1978 Jul;75(7):3479–3483. doi: 10.1073/pnas.75.7.3479. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Dunny G. M., Craig R. A., Carron R. L., Clewell D. B. Plasmid transfer in Streptococcus faecalis: production of multiple sex pheromones by recipients. Plasmid. 1979 Jul;2(3):454–465. doi: 10.1016/0147-619x(79)90029-5. [DOI] [PubMed] [Google Scholar]
  8. Dunny G. M., Zimmerman D. L., Tortorello M. L. Induction of surface exclusion (entry exclusion) by Streptococcus faecalis sex pheromones: use of monoclonal antibodies to identify an inducible surface antigen involved in the exclusion process. Proc Natl Acad Sci U S A. 1985 Dec;82(24):8582–8586. doi: 10.1073/pnas.82.24.8582. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Ehrenfeld E. E., Clewell D. B. Transfer functions of the Streptococcus faecalis plasmid pAD1: organization of plasmid DNA encoding response to sex pheromone. J Bacteriol. 1987 Aug;169(8):3473–3481. doi: 10.1128/jb.169.8.3473-3481.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Ehrenfeld E. E., Kessler R. E., Clewell D. B. Identification of pheromone-induced surface proteins in Streptococcus faecalis and evidence of a role for lipoteichoic acid in formation of mating aggregates. J Bacteriol. 1986 Oct;168(1):6–12. doi: 10.1128/jb.168.1.6-12.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Green P. J., Pines O., Inouye M. The role of antisense RNA in gene regulation. Annu Rev Biochem. 1986;55:569–597. doi: 10.1146/annurev.bi.55.070186.003033. [DOI] [PubMed] [Google Scholar]
  12. Gryczan T. J., Hahn J., Contente S., Dubnau D. Replication and incompatibility properties of plasmid pE194 in Bacillus subtilis. J Bacteriol. 1982 Nov;152(2):722–735. doi: 10.1128/jb.152.2.722-735.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Ike Y., Clewell D. B. Genetic analysis of the pAD1 pheromone response in Streptococcus faecalis, using transposon Tn917 as an insertional mutagen. J Bacteriol. 1984 Jun;158(3):777–783. doi: 10.1128/jb.158.3.777-783.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Ike Y., Craig R. A., White B. A., Yagi Y., Clewell D. B. Modification of Streptococcus faecalis sex pheromones after acquisition of plasmid DNA. Proc Natl Acad Sci U S A. 1983 Sep;80(17):5369–5373. doi: 10.1073/pnas.80.17.5369. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Kessler R. E., Yagi Y. Identification and partial characterization of a pheromone-induced adhesive surface antigen of Streptococcus faecalis. J Bacteriol. 1983 Aug;155(2):714–721. doi: 10.1128/jb.155.2.714-721.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  17. Mori M., Sakagami Y., Narita M., Isogai A., Fujino M., Kitada C., Craig R. A., Clewell D. B., Suzuki A. Isolation and structure of the bacterial sex pheromone, cAD1, that induces plasmid transfer in Streptococcus faecalis. FEBS Lett. 1984 Dec 3;178(1):97–100. doi: 10.1016/0014-5793(84)81248-x. [DOI] [PubMed] [Google Scholar]
  18. Perkins J. B., Youngman P. J. Construction and properties of Tn917-lac, a transposon derivative that mediates transcriptional gene fusions in Bacillus subtilis. Proc Natl Acad Sci U S A. 1986 Jan;83(1):140–144. doi: 10.1073/pnas.83.1.140. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Shaw J. H., Clewell D. B. Complete nucleotide sequence of macrolide-lincosamide-streptogramin B-resistance transposon Tn917 in Streptococcus faecalis. J Bacteriol. 1985 Nov;164(2):782–796. doi: 10.1128/jb.164.2.782-796.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Suzuki A., Mori M., Sakagami Y., Isogai A., Fujino M., Kitada C., Craig R. A., Clewell D. B. Isolation and structure of bacterial sex pheromone, cPD1. Science. 1984 Nov 16;226(4676):849–850. doi: 10.1126/science.6436978. [DOI] [PubMed] [Google Scholar]
  21. Tomich P. K., An F. Y., Clewell D. B. Properties of erythromycin-inducible transposon Tn917 in Streptococcus faecalis. J Bacteriol. 1980 Mar;141(3):1366–1374. doi: 10.1128/jb.141.3.1366-1374.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Tomich P. K., An F. Y., Damle S. P., Clewell D. B. Plasmid-related transmissibility and multiple drug resistance in Streptococcus faecalis subsp. zymogenes strain DS16. Antimicrob Agents Chemother. 1979 Jun;15(6):828–830. doi: 10.1128/aac.15.6.828. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Tortorello M. L., Dunny G. M. Identification of multiple cell surface antigens associated with the sex pheromone response of Streptococcus faecalis. J Bacteriol. 1985 Apr;162(1):131–137. doi: 10.1128/jb.162.1.131-137.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Towbin H., Staehelin T., Gordon J. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci U S A. 1979 Sep;76(9):4350–4354. doi: 10.1073/pnas.76.9.4350. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Wirth R., An F. Y., Clewell D. B. Highly efficient protoplast transformation system for Streptococcus faecalis and a new Escherichia coli-S. faecalis shuttle vector. J Bacteriol. 1986 Mar;165(3):831–836. doi: 10.1128/jb.165.3.831-836.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Yagi Y., Kessler R. E., Shaw J. H., Lopatin D. E., An F., Clewell D. B. Plasmid content of Streptococcus faecalis strain 39-5 and identification of a pheromone (cPD1)-induced surface antigen. J Gen Microbiol. 1983 Apr;129(4):1207–1215. doi: 10.1099/00221287-129-4-1207. [DOI] [PubMed] [Google Scholar]

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