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. 1996 Aug;178(15):4563–4570. doi: 10.1128/jb.178.15.4563-4570.1996

The molecular architecture of the sar locus in Staphylococcus aureus.

M G Bayer 1, J H Heinrichs 1, A L Cheung 1
PMCID: PMC178224  PMID: 8755885

Abstract

The global regulator sar in Staphylococcus aureus controls the synthesis of a variety of cell wall and extracellular proteins, many of which are putative virulence factors. The sar locus in strain RN6390 contains a 339-bp open reading frame (sarA) and an 860-bp upstream region. Transcriptional analyses of this locus revealed three different transcripts of 0.58, 0.84, and 1.15 kb (designated sarA, sarC, and sarB, respectively). All three transcripts seemed to be under temporal, growth cycle-dependent regulation, with sarA and sarB being most abundant in early log phase and the sarC concentration being highest toward the late stationary phase. Mapping of the 5' ends of the sar transcripts by primer extension and modified S1 nuclease protection assays demonstrated that transcription is initiated from three separate, widely spaced promoters. The 3' ends of all three sar transcripts are identical, and transcriptional termination occurs upstream of a typical prokaryotic poly(T) termination signal. Northern (RNA) analysis of sar mutant clones containing plasmids that comprised various promoters and the termination signal revealed that individual transcripts can be generated from each of the three promoters, thus suggesting possible activation as independent promoters. The multipromoter system, from which transcription is initiated, bears conserved features for recognition by homologous sigma 70 transcription factors and also by those expressed in the general stress response. Downstream of the two distal promoters (P3 and P2) are two regions potentially encoding short peptides. It is conceivable that posttranslational cooperation between these short peptides and the sarA gene product occurs to modulate sar-related functions. Complementation studies of a sar mutant with a clone expressing all three sar transcripts showed that this clone was able to restore the sar wild-type phenotype to the sar mutant.

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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. Boylan S. A., Thomas M. D., Price C. W. Genetic method to identify regulons controlled by nonessential elements: isolation of a gene dependent on alternate transcription factor sigma B of Bacillus subtilis. J Bacteriol. 1991 Dec;173(24):7856–7866. doi: 10.1128/jb.173.24.7856-7866.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Brendel V., Hamm G. H., Trifonov E. N. Terminators of transcription with RNA polymerase from Escherichia coli: what they look like and how to find them. J Biomol Struct Dyn. 1986 Feb;3(4):705–723. doi: 10.1080/07391102.1986.10508457. [DOI] [PubMed] [Google Scholar]
  4. Brendel V., Trifonov E. N. A computer algorithm for testing potential prokaryotic terminators. Nucleic Acids Res. 1984 May 25;12(10):4411–4427. doi: 10.1093/nar/12.10.4411. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Chang S., Cohen S. N. High frequency transformation of Bacillus subtilis protoplasts by plasmid DNA. Mol Gen Genet. 1979 Jan 5;168(1):111–115. doi: 10.1007/BF00267940. [DOI] [PubMed] [Google Scholar]
  6. 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]
  7. 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]
  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., 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]
  11. 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]
  12. Greene C., McDevitt D., Francois P., Vaudaux P. E., Lew D. P., Foster T. J. Adhesion properties of mutants of Staphylococcus aureus defective in fibronectin-binding proteins and studies on the expression of fnb genes. Mol Microbiol. 1995 Sep;17(6):1143–1152. doi: 10.1111/j.1365-2958.1995.mmi_17061143.x. [DOI] [PubMed] [Google Scholar]
  13. Heinrichs J. H., Bayer M. G., Cheung A. L. Characterization of the sar locus and its interaction with agr in Staphylococcus aureus. J Bacteriol. 1996 Jan;178(2):418–423. doi: 10.1128/jb.178.2.418-423.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Helmann J. D. Compilation and analysis of Bacillus subtilis sigma A-dependent promoter sequences: evidence for extended contact between RNA polymerase and upstream promoter DNA. Nucleic Acids Res. 1995 Jul 11;23(13):2351–2360. doi: 10.1093/nar/23.13.2351. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Janzon L., Arvidson S. The role of the delta-lysin gene (hld) in the regulation of virulence genes by the accessory gene regulator (agr) in Staphylococcus aureus. EMBO J. 1990 May;9(5):1391–1399. doi: 10.1002/j.1460-2075.1990.tb08254.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Jeon Y. H., Negishi T., Shirakawa M., Yamazaki T., Fujita N., Ishihama A., Kyogoku Y. Solution structure of the activator contact domain of the RNA polymerase alpha subunit. Science. 1995 Dec 1;270(5241):1495–1497. doi: 10.1126/science.270.5241.1495. [DOI] [PubMed] [Google Scholar]
  17. Liu-Johnson H. N., Gartenberg M. R., Crothers D. M. The DNA binding domain and bending angle of E. coli CAP protein. Cell. 1986 Dec 26;47(6):995–1005. doi: 10.1016/0092-8674(86)90814-7. [DOI] [PubMed] [Google Scholar]
  18. Mager W. H., De Kruijff A. J. Stress-induced transcriptional activation. Microbiol Rev. 1995 Sep;59(3):506–531. doi: 10.1128/mr.59.3.506-531.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Mager W. H., De Kruijff A. J. Stress-induced transcriptional activation. Microbiol Rev. 1995 Sep;59(3):506–531. doi: 10.1128/mr.59.3.506-531.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Miksch G., Dobrowolski P. Growth phase-dependent induction of stationary-phase promoters of Escherichia coli in different gram-negative bacteria. J Bacteriol. 1995 Sep;177(18):5374–5378. doi: 10.1128/jb.177.18.5374-5378.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. 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]
  22. Pérez-Martín J., Rojo F., de Lorenzo V. Promoters responsive to DNA bending: a common theme in prokaryotic gene expression. Microbiol Rev. 1994 Jun;58(2):268–290. doi: 10.1128/mr.58.2.268-290.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Rao L., Karls R. K., Betley M. J. In vitro transcription of pathogenesis-related genes by purified RNA polymerase from Staphylococcus aureus. J Bacteriol. 1995 May;177(10):2609–2614. doi: 10.1128/jb.177.10.2609-2614.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Ross W., Gosink K. K., Salomon J., Igarashi K., Zou C., Ishihama A., Severinov K., Gourse R. L. A third recognition element in bacterial promoters: DNA binding by the alpha subunit of RNA polymerase. Science. 1993 Nov 26;262(5138):1407–1413. doi: 10.1126/science.8248780. [DOI] [PubMed] [Google Scholar]
  25. Vicente M., Kushner S. R., Garrido T., Aldea M. The role of the 'gearbox' in the transcription of essential genes. Mol Microbiol. 1991 Sep;5(9):2085–2091. doi: 10.1111/j.1365-2958.1991.tb02137.x. [DOI] [PubMed] [Google Scholar]

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