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. 1995 Jun;177(11):3220–3226. doi: 10.1128/jb.177.11.3220-3226.1995

Insertional inactivation of a chromosomal locus that modulates expression of potential virulence determinants in Staphylococcus aureus.

A L Cheung 1, C Wolz 1, M R Yeaman 1, A S Bayer 1
PMCID: PMC177014  PMID: 7539418

Abstract

A single insertion of transposon Tn551 into a unique chromosomal locus of Staphylococcus aureus ISP479C has resulted in a pleiotropic effect on the expression of both extracellular and cell wall proteins. In particular, the expression of cell wall protein A and clumping activity with fibrinogen were rendered undetectable in the mutant 1E3 compared with the parent. The secretion of alpha-hemolysin in mutant 1E3 was modestly increased. Southern blot and phenotypic analyses indicated that this locus is distinct from agr, xpr, and sar, three previously described global regulatory loci. Transduction experiments demonstrated that the genotype associated with mutant 1E3 could be transferred back into the parental strain ISP479C. The transductant 1E3-2 displayed a phenotypic profile similar to that of the original mutant. Northern (RNA) blot studies showed that this locus may be involved in modulating target genes at the mRNA level. In the rabbit endocarditis model, there was a significant decrease in both the infectivity rate and intravegetation bacterial density with mutant 1E3 compared with the parent at an inoculum of 10(3) CFU. Since protein A and the fibrinogen-binding protein(s) are major surface proteins that may mediate bacterial adhesion to host tissues, this locus may be an important genetic element involved in the expression of virulence determinants in S. aureus.

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Selected References

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  1. Blake M. S., Johnston K. H., Russell-Jones G. J., Gotschlich E. C. A rapid, sensitive method for detection of alkaline phosphatase-conjugated anti-antibody on Western blots. Anal Biochem. 1984 Jan;136(1):175–179. doi: 10.1016/0003-2697(84)90320-8. [DOI] [PubMed] [Google Scholar]
  2. Bodén M. K., Flock J. I. Cloning and characterization of a gene for a 19 kDa fibrinogen-binding protein from Staphylococcus aureus. Mol Microbiol. 1994 May;12(4):599–606. doi: 10.1111/j.1365-2958.1994.tb01046.x. [DOI] [PubMed] [Google Scholar]
  3. Camilli A., Portnoy A., Youngman P. Insertional mutagenesis of Listeria monocytogenes with a novel Tn917 derivative that allows direct cloning of DNA flanking transposon insertions. J Bacteriol. 1990 Jul;172(7):3738–3744. doi: 10.1128/jb.172.7.3738-3744.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Cheung A. I., Projan S. J., Edelstein R. E., Fischetti V. A. Cloning, expression, and nucleotide sequence of a Staphylococcus aureus gene (fbpA) encoding a fibrinogen-binding protein. Infect Immun. 1995 May;63(5):1914–1920. doi: 10.1128/iai.63.5.1914-1920.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Cheung A. L., Eberhardt K. J., Chung E., Yeaman M. R., Sullam P. M., Ramos M., Bayer A. S. Diminished virulence of a sar-/agr- mutant of Staphylococcus aureus in the rabbit model of endocarditis. J Clin Invest. 1994 Nov;94(5):1815–1822. doi: 10.1172/JCI117530. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Cheung A. L., Eberhardt K. J., Fischetti V. A. A method to isolate RNA from gram-positive bacteria and mycobacteria. Anal Biochem. 1994 Nov 1;222(2):511–514. doi: 10.1006/abio.1994.1528. [DOI] [PubMed] [Google Scholar]
  7. Cheung A. L., Fischetti V. A. The role of fibrinogen in staphylococcal adherence to catheters in vitro. J Infect Dis. 1990 Jun;161(6):1177–1186. doi: 10.1093/infdis/161.6.1177. [DOI] [PubMed] [Google Scholar]
  8. Cheung A. L., Fischetti V. A. Variation in the expression of cell wall proteins of Staphylococcus aureus grown on solid and liquid media. Infect Immun. 1988 May;56(5):1061–1065. doi: 10.1128/iai.56.5.1061-1065.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Cheung A. L., Koomey J. M., Butler C. A., Projan S. J., Fischetti V. A. Regulation of exoprotein expression in Staphylococcus aureus by a locus (sar) distinct from agr. Proc Natl Acad Sci U S A. 1992 Jul 15;89(14):6462–6466. doi: 10.1073/pnas.89.14.6462. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Cheung A. L., Krishnan M., Jaffe E. A., Fischetti V. A. Fibrinogen acts as a bridging molecule in the adherence of Staphylococcus aureus to cultured human endothelial cells. J Clin Invest. 1991 Jun;87(6):2236–2245. doi: 10.1172/JCI115259. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Cheung A. L., Projan S. J. Cloning and sequencing of sarA of Staphylococcus aureus, a gene required for the expression of agr. J Bacteriol. 1994 Jul;176(13):4168–4172. doi: 10.1128/jb.176.13.4168-4172.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Cheung A. L., Yeaman M. R., Sullam P. M., Witt M. D., Bayer A. S. Role of the sar locus of Staphylococcus aureus in induction of endocarditis in rabbits. Infect Immun. 1994 May;62(5):1719–1725. doi: 10.1128/iai.62.5.1719-1725.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Cheung A. L., Ying P. Regulation of alpha- and beta-hemolysins by the sar locus of Staphylococcus aureus. J Bacteriol. 1994 Feb;176(3):580–585. doi: 10.1128/jb.176.3.580-585.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Durack D. T., Beeson P. B. Experimental bacterial endocarditis. I. Colonization of a sterile vegetation. Br J Exp Pathol. 1972 Feb;53(1):44–49. [PMC free article] [PubMed] [Google Scholar]
  15. Fröman G., Switalski L. M., Speziale P., Hök M. Isolation and characterization of a fibronectin receptor from Staphylococcus aureus. J Biol Chem. 1987 May 15;262(14):6564–6571. [PubMed] [Google Scholar]
  16. Hart M. E., Smeltzer M. S., Iandolo J. J. The extracellular protein regulator (xpr) affects exoprotein and agr mRNA levels in Staphylococcus aureus. J Bacteriol. 1993 Dec;175(24):7875–7879. doi: 10.1128/jb.175.24.7875-7879.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Laddaga R. A., Chu L., Misra T. K., Silver S. Nucleotide sequence and expression of the mercurial-resistance operon from Staphylococcus aureus plasmid pI258. Proc Natl Acad Sci U S A. 1987 Aug;84(15):5106–5110. doi: 10.1073/pnas.84.15.5106. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. 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]
  19. Lantz M. S., Allen R. D., Bounelis P., Switalski L. M., Hook M. Bacteroides gingivalis and Bacteroides intermedius recognize different sites on human fibrinogen. J Bacteriol. 1990 Feb;172(2):716–726. doi: 10.1128/jb.172.2.716-726.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. McDevitt D., Francois P., Vaudaux P., Foster T. J. Molecular characterization of the clumping factor (fibrinogen receptor) of Staphylococcus aureus. Mol Microbiol. 1994 Jan;11(2):237–248. doi: 10.1111/j.1365-2958.1994.tb00304.x. [DOI] [PubMed] [Google Scholar]
  21. Nixon B. T., Ronson C. W., Ausubel F. M. Two-component regulatory systems responsive to environmental stimuli share strongly conserved domains with the nitrogen assimilation regulatory genes ntrB and ntrC. Proc Natl Acad Sci U S A. 1986 Oct;83(20):7850–7854. doi: 10.1073/pnas.83.20.7850. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Novick R. P., Adler G. K., Projan S. J., Carleton S., Highlander S. K., Gruss A., Khan S. A., Iordanescu S. Control of pT181 replication I. The pT181 copy control function acts by inhibiting the synthesis of a replication protein. EMBO J. 1984 Oct;3(10):2399–2405. doi: 10.1002/j.1460-2075.1984.tb02146.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Novick R. P., Ross H. F., Projan S. J., Kornblum J., Kreiswirth B., Moghazeh S. Synthesis of staphylococcal virulence factors is controlled by a regulatory RNA molecule. EMBO J. 1993 Oct;12(10):3967–3975. doi: 10.1002/j.1460-2075.1993.tb06074.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. O'Reilly M., de Azavedo J. C., Kennedy S., Foster T. J. Inactivation of the alpha-haemolysin gene of Staphylococcus aureus 8325-4 by site-directed mutagenesis and studies on the expression of its haemolysins. Microb Pathog. 1986 Apr;1(2):125–138. doi: 10.1016/0882-4010(86)90015-x. [DOI] [PubMed] [Google Scholar]
  25. Patel A. H., Kornblum J., Kreiswirth B., Novick R., Foster T. J. Regulation of the protein A-encoding gene in Staphylococcus aureus. Gene. 1992 May 1;114(1):25–34. doi: 10.1016/0378-1119(92)90703-r. [DOI] [PubMed] [Google Scholar]
  26. Patel A. H., Nowlan P., Weavers E. D., Foster T. Virulence of protein A-deficient and alpha-toxin-deficient mutants of Staphylococcus aureus isolated by allele replacement. Infect Immun. 1987 Dec;55(12):3103–3110. doi: 10.1128/iai.55.12.3103-3110.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Peterson P. K., Verhoef J., Sabath L. D., Quie P. G. Effect of protein A on staphylococcal opsonization. Infect Immun. 1977 Mar;15(3):760–764. doi: 10.1128/iai.15.3.760-764.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Recsei P., Kreiswirth B., O'Reilly M., Schlievert P., Gruss A., Novick R. P. Regulation of exoprotein gene expression in Staphylococcus aureus by agar. Mol Gen Genet. 1986 Jan;202(1):58–61. doi: 10.1007/BF00330517. [DOI] [PubMed] [Google Scholar]
  29. Smeltzer M. S., Gill S. R., Iandolo J. J. Localization of a chromosomal mutation affecting expression of extracellular lipase in Staphylococcus aureus. J Bacteriol. 1992 Jun;174(12):4000–4006. doi: 10.1128/jb.174.12.4000-4006.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Smeltzer M. S., Hart M. E., Iandolo J. J. Phenotypic characterization of xpr, a global regulator of extracellular virulence factors in Staphylococcus aureus. Infect Immun. 1993 Mar;61(3):919–925. doi: 10.1128/iai.61.3.919-925.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. 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]
  32. Vandenesch F., Kornblum J., Novick R. P. A temporal signal, independent of agr, is required for hla but not spa transcription in Staphylococcus aureus. J Bacteriol. 1991 Oct;173(20):6313–6320. doi: 10.1128/jb.173.20.6313-6320.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]

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