Abstract
To determine whether growth phase affects the expression of mga and other virulence-associated genes in the group A streptococcus (GAS), total RNA was isolated from the serotype M6 GAS strain JRS4 at different phases of growth and transcript levels were quantitated by hybridization with radiolabeled DNA probes. Expression of mga (which encodes a multiple gene regulator) and the Mga-regulated genes emm (which encodes M protein) and scpA (which encodes a complement C5a peptidase) was found to be maximal in exponential phase and shut off as the bacteria entered stationary phase, while the housekeeping genes recA and rpsL showed constant transcript levels over the same period of growth. Expression of mga from a foreign phage promoter in a mga-deleted GAS strain (JRS519) altered the wild-type growth phase-dependent transcription profile seen for emm and scpA, as well as for mga. Therefore, the temporal control of mga expression requires its upstream promoter region, and the subsequent growth phase regulation of emm and scpA is Mga dependent. A number of putative virulence genes in JRS4 were shown not to require Mga for their expression, although several exhibited growth phase-dependent regulation that was similar to mga, i.e., slo (which encodes streptolysin O) and plr (encoding the plasmin receptor/glyceraldehyde-3-phosphate dehydrogenase). Still others showed a markedly different pattern of expression (the genes for the superantigen toxins MF and SpeC). These results suggest the existence of complex levels of global regulation sensitive to growth phase that directly control the expression of virulence genes and mga in GAS.
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- Andersson G., McIver K., Hedén L. O., Scott J. R. Complementation of divergent mga genes in group A Streptococcus. Gene. 1996 Oct 10;175(1-2):77–81. doi: 10.1016/0378-1119(96)00124-2. [DOI] [PubMed] [Google Scholar]
- Balaban N., Novick R. P. Autocrine regulation of toxin synthesis by Staphylococcus aureus. Proc Natl Acad Sci U S A. 1995 Feb 28;92(5):1619–1623. doi: 10.1073/pnas.92.5.1619. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Benson A. K., Haldenwang W. G. Characterization of a regulatory network that controls sigma B expression in Bacillus subtilis. J Bacteriol. 1992 Feb;174(3):749–757. doi: 10.1128/jb.174.3.749-757.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Bisno A. L., Stevens D. L. Streptococcal infections of skin and soft tissues. N Engl J Med. 1996 Jan 25;334(4):240–245. doi: 10.1056/NEJM199601253340407. [DOI] [PubMed] [Google Scholar]
- Boylan S. A., Rutherford A., Thomas S. M., Price C. W. Activation of Bacillus subtilis transcription factor sigma B by a regulatory pathway responsive to stationary-phase signals. J Bacteriol. 1992 Jun;174(11):3695–3706. doi: 10.1128/jb.174.11.3695-3706.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Caparon M. G., Geist R. T., Perez-Casal J., Scott J. R. Environmental regulation of virulence in group A streptococci: transcription of the gene encoding M protein is stimulated by carbon dioxide. J Bacteriol. 1992 Sep;174(17):5693–5701. doi: 10.1128/jb.174.17.5693-5701.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Caparon M. G., Scott J. R. Excision and insertion of the conjugative transposon Tn916 involves a novel recombination mechanism. Cell. 1989 Dec 22;59(6):1027–1034. doi: 10.1016/0092-8674(89)90759-9. [DOI] [PubMed] [Google Scholar]
- Caparon M. G., Scott J. R. Identification of a gene that regulates expression of M protein, the major virulence determinant of group A streptococci. Proc Natl Acad Sci U S A. 1987 Dec;84(23):8677–8681. doi: 10.1073/pnas.84.23.8677. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Chassy B. M. A gentle method for the lysis of oral streptococci. Biochem Biophys Res Commun. 1976 Jan 26;68(2):603–608. doi: 10.1016/0006-291x(76)91188-8. [DOI] [PubMed] [Google Scholar]
- Chen C., Bormann N., Cleary P. P. VirR and Mry are homologous trans-acting regulators of M protein and C5a peptidase expression in group A streptococci. Mol Gen Genet. 1993 Dec;241(5-6):685–693. doi: 10.1007/BF00279912. [DOI] [PubMed] [Google Scholar]
- 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]
- Cleary P., Retnoningrum D. Group A streptococcal immunoglobulin-binding proteins: adhesins, molecular mimicry or sensory proteins? Trends Microbiol. 1994 Apr;2(4):131–136. doi: 10.1016/0966-842x(94)90600-9. [DOI] [PubMed] [Google Scholar]
- Coote J. G. Antigenic switching and pathogenicity: environmental effects on virulence gene expression in Bordetella pertussis. J Gen Microbiol. 1991 Nov;137(11):2493–2503. doi: 10.1099/00221287-137-11-2493. [DOI] [PubMed] [Google Scholar]
- Crater D. L., van de Rijn I. Hyaluronic acid synthesis operon (has) expression in group A streptococci. J Biol Chem. 1995 Aug 4;270(31):18452–18458. doi: 10.1074/jbc.270.31.18452. [DOI] [PubMed] [Google Scholar]
- Eichenbaum Z., Green B. D., Scott J. R. Iron starvation causes release from the group A streptococcus of the ADP-ribosylating protein called plasmin receptor or surface glyceraldehyde-3-phosphate-dehydrogenase. Infect Immun. 1996 Jun;64(6):1956–1960. doi: 10.1128/iai.64.6.1956-1960.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Glisin V., Crkvenjakov R., Byus C. Ribonucleic acid isolated by cesium chloride centrifugation. Biochemistry. 1974 Jun 4;13(12):2633–2637. doi: 10.1021/bi00709a025. [DOI] [PubMed] [Google Scholar]
- Grossman A. D. Genetic networks controlling the initiation of sporulation and the development of genetic competence in Bacillus subtilis. Annu Rev Genet. 1995;29:477–508. doi: 10.1146/annurev.ge.29.120195.002401. [DOI] [PubMed] [Google Scholar]
- Gunn J. S., Miller S. I. PhoP-PhoQ activates transcription of pmrAB, encoding a two-component regulatory system involved in Salmonella typhimurium antimicrobial peptide resistance. J Bacteriol. 1996 Dec;178(23):6857–6864. doi: 10.1128/jb.178.23.6857-6864.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Hanahan D. Studies on transformation of Escherichia coli with plasmids. J Mol Biol. 1983 Jun 5;166(4):557–580. doi: 10.1016/s0022-2836(83)80284-8. [DOI] [PubMed] [Google Scholar]
- Hauser A. R., Stevens D. L., Kaplan E. L., Schlievert P. M. Molecular analysis of pyrogenic exotoxins from Streptococcus pyogenes isolates associated with toxic shock-like syndrome. J Clin Microbiol. 1991 Aug;29(8):1562–1567. doi: 10.1128/jcm.29.8.1562-1567.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Hoge C. W., Schwartz B., Talkington D. F., Breiman R. F., MacNeill E. M., Englender S. J. The changing epidemiology of invasive group A streptococcal infections and the emergence of streptococcal toxic shock-like syndrome. A retrospective population-based study. JAMA. 1993 Jan 20;269(3):384–389. [PubMed] [Google Scholar]
- Husmann L. K., Scott J. R., Lindahl G., Stenberg L. Expression of the Arp protein, a member of the M protein family, is not sufficient to inhibit phagocytosis of Streptococcus pyogenes. Infect Immun. 1995 Jan;63(1):345–348. doi: 10.1128/iai.63.1.345-348.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Kihlberg B. M., Cooney J., Caparon M. G., Olsén A., Björck L. Biological properties of a Streptococcus pyogenes mutant generated by Tn916 insertion in mga. Microb Pathog. 1995 Nov;19(5):299–315. doi: 10.1016/s0882-4010(96)80003-9. [DOI] [PubMed] [Google Scholar]
- LANCEFIELD R. C. Current knowledge of type-specific M antigens of group A streptococci. J Immunol. 1962 Sep;89:307–313. [PubMed] [Google Scholar]
- Lebeau C., Vandenesch F., Greenland T., Novick R. P., Etienne J. Coagulase expression in Staphylococcus aureus is positively and negatively modulated by an agr-dependent mechanism. J Bacteriol. 1994 Sep;176(17):5534–5536. doi: 10.1128/jb.176.17.5534-5536.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Loewen P. C., Hengge-Aronis R. The role of the sigma factor sigma S (KatF) in bacterial global regulation. Annu Rev Microbiol. 1994;48:53–80. doi: 10.1146/annurev.mi.48.100194.000413. [DOI] [PubMed] [Google Scholar]
- Lottenberg R., Broder C. C., Boyle M. D., Kain S. J., Schroeder B. L., Curtiss R., 3rd Cloning, sequence analysis, and expression in Escherichia coli of a streptococcal plasmin receptor. J Bacteriol. 1992 Aug;174(16):5204–5210. doi: 10.1128/jb.174.16.5204-5210.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Macdonald P. M., Kutter E., Mosig G. Regulation of a bacteriophage T4 late gene, soc, which maps in an early region. Genetics. 1984 Jan;106(1):17–27. doi: 10.1093/genetics/106.1.17. [DOI] [PMC free article] [PubMed] [Google Scholar]
- McIver K. S., Heath A. S., Scott J. R. Regulation of virulence by environmental signals in group A streptococci: influence of osmolarity, temperature, gas exchange, and iron limitation on emm transcription. Infect Immun. 1995 Nov;63(11):4540–4542. doi: 10.1128/iai.63.11.4540-4542.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
- McLandsborough L. A., Cleary P. P. Insertional inactivation of virR in Streptococcus pyogenes M49 demonstrates that VirR functions as a positive regulator of streptococcal C5a peptidase and M protein in OF+ strains. Dev Biol Stand. 1995;85:149–152. [PubMed] [Google Scholar]
- Morfeldt E., Janzon L., Arvidson S., Löfdahl S. Cloning of a chromosomal locus (exp) which regulates the expression of several exoprotein genes in Staphylococcus aureus. Mol Gen Genet. 1988 Mar;211(3):435–440. doi: 10.1007/BF00425697. [DOI] [PubMed] [Google Scholar]
- Novotny W., Faden H., Mosovich L. Emergence of invasive group A streptococcal disease among young children. Clin Pediatr (Phila) 1992 Oct;31(10):596–601. doi: 10.1177/000992289203101004. [DOI] [PubMed] [Google Scholar]
- O'Reilly M., Devine K. M. Expression of AbrB, a transition state regulator from Bacillus subtilis, is growth phase dependent in a manner resembling that of Fis, the nucleoid binding protein from Escherichia coli. J Bacteriol. 1997 Jan;179(2):522–529. doi: 10.1128/jb.179.2.522-529.1997. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Okada N., Geist R. T., Caparon M. G. Positive transcriptional control of mry regulates virulence in the group A streptococcus. Mol Microbiol. 1993 Mar;7(6):893–903. doi: 10.1111/j.1365-2958.1993.tb01180.x. [DOI] [PubMed] [Google Scholar]
- Otto R., de Vos W. M., Gavrieli J. Plasmid DNA in Streptococcus cremoris Wg2: Influence of pH on Selection in Chemostats of a Variant Lacking a Protease Plasmid. Appl Environ Microbiol. 1982 Jun;43(6):1272–1277. doi: 10.1128/aem.43.6.1272-1277.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Pancholi V., Fischetti V. A. A major surface protein on group A streptococci is a glyceraldehyde-3-phosphate-dehydrogenase with multiple binding activity. J Exp Med. 1992 Aug 1;176(2):415–426. doi: 10.1084/jem.176.2.415. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Peng H. L., Novick R. P., Kreiswirth B., Kornblum J., Schlievert P. Cloning, characterization, and sequencing of an accessory gene regulator (agr) in Staphylococcus aureus. J Bacteriol. 1988 Sep;170(9):4365–4372. doi: 10.1128/jb.170.9.4365-4372.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Perez-Casal J., Caparon M. G., Scott J. R. Mry, a trans-acting positive regulator of the M protein gene of Streptococcus pyogenes with similarity to the receptor proteins of two-component regulatory systems. J Bacteriol. 1991 Apr;173(8):2617–2624. doi: 10.1128/jb.173.8.2617-2624.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Perez-Casal J., Price J. A., Maguin E., Scott J. R. An M protein with a single C repeat prevents phagocytosis of Streptococcus pyogenes: use of a temperature-sensitive shuttle vector to deliver homologous sequences to the chromosome of S. pyogenes. Mol Microbiol. 1993 May;8(5):809–819. doi: 10.1111/j.1365-2958.1993.tb01628.x. [DOI] [PubMed] [Google Scholar]
- Pestova E. V., Håvarstein L. S., Morrison D. A. Regulation of competence for genetic transformation in Streptococcus pneumoniae by an auto-induced peptide pheromone and a two-component regulatory system. Mol Microbiol. 1996 Aug;21(4):853–862. doi: 10.1046/j.1365-2958.1996.501417.x. [DOI] [PubMed] [Google Scholar]
- Platteeuw C., Simons G., de Vos W. M. Use of the Escherichia coli beta-glucuronidase (gusA) gene as a reporter gene for analyzing promoters in lactic acid bacteria. Appl Environ Microbiol. 1994 Feb;60(2):587–593. doi: 10.1128/aem.60.2.587-593.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Podbielski A., Flosdorff A., Weber-Heynemann J. The group A streptococcal virR49 gene controls expression of four structural vir regulon genes. Infect Immun. 1995 Jan;63(1):9–20. doi: 10.1128/iai.63.1.9-20.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Podbielski A., Peterson J. A., Cleary P. Surface protein-CAT reporter fusions demonstrate differential gene expression in the vir regulon of Streptococcus pyogenes. Mol Microbiol. 1992 Aug;6(16):2253–2265. doi: 10.1111/j.1365-2958.1992.tb01401.x. [DOI] [PubMed] [Google Scholar]
- 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]
- Sanger F., Nicklen S., Coulson A. R. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. doi: 10.1073/pnas.74.12.5463. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Scarlato V., Aricò B., Goyard S., Ricci S., Manetti R., Prugnola A., Manetti R., Polverino-De-Laureto P., Ullmann A., Rappuoli R. A novel chromatin-forming histone H1 homologue is encoded by a dispensable and growth-regulated gene in Bordetella pertussis. Mol Microbiol. 1995 Mar;15(5):871–881. doi: 10.1111/j.1365-2958.1995.tb02357.x. [DOI] [PubMed] [Google Scholar]
- Schlievert P. M. Role of superantigens in human disease. J Infect Dis. 1993 May;167(5):997–1002. doi: 10.1093/infdis/167.5.997. [DOI] [PubMed] [Google Scholar]
- Scott J. R. A new gene controlling lysogeny in phage P1. Virology. 1972 Apr;48(1):282–283. doi: 10.1016/0042-6822(72)90139-0. [DOI] [PubMed] [Google Scholar]
- Scott J. R., Guenthner P. C., Malone L. M., Fischetti V. A. Conversion of an M- group A streptococcus to M+ by transfer of a plasmid containing an M6 gene. J Exp Med. 1986 Nov 1;164(5):1641–1651. doi: 10.1084/jem.164.5.1641. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 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]
- Soncini F. C., Groisman E. A. Two-component regulatory systems can interact to process multiple environmental signals. J Bacteriol. 1996 Dec;178(23):6796–6801. doi: 10.1128/jb.178.23.6796-6801.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Steffen P., Goyard S., Ullmann A. Phosphorylated BvgA is sufficient for transcriptional activation of virulence-regulated genes in Bordetella pertussis. EMBO J. 1996 Jan 2;15(1):102–109. [PMC free article] [PubMed] [Google Scholar]
- Stenberg L., O'Toole P., Lindahl G. Many group A streptococcal strains express two different immunoglobulin-binding proteins, encoded by closely linked genes: characterization of the proteins expressed by four strains of different M-type. Mol Microbiol. 1992 May;6(9):1185–1194. doi: 10.1111/j.1365-2958.1992.tb01557.x. [DOI] [PubMed] [Google Scholar]
- Stevens D. L. Invasive group A streptococcus infections. Clin Infect Dis. 1992 Jan;14(1):2–11. doi: 10.1093/clinids/14.1.2. [DOI] [PubMed] [Google Scholar]
- Stevens D. L. Streptococcal toxic-shock syndrome: spectrum of disease, pathogenesis, and new concepts in treatment. Emerg Infect Dis. 1995 Jul-Sep;1(3):69–78. doi: 10.3201/eid0103.950301. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Strauch M. A., Hoch J. A. Transition-state regulators: sentinels of Bacillus subtilis post-exponential gene expression. Mol Microbiol. 1993 Feb;7(3):337–342. doi: 10.1111/j.1365-2958.1993.tb01125.x. [DOI] [PubMed] [Google Scholar]
- Thomas P. S. Hybridization of denatured RNA and small DNA fragments transferred to nitrocellulose. Proc Natl Acad Sci U S A. 1980 Sep;77(9):5201–5205. doi: 10.1073/pnas.77.9.5201. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 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]
- Wexler D. E., Chenoweth D. E., Cleary P. P. Mechanism of action of the group A streptococcal C5a inactivator. Proc Natl Acad Sci U S A. 1985 Dec;82(23):8144–8148. doi: 10.1073/pnas.82.23.8144. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Wexler D. E., Cleary P. P. Purification and characteristics of the streptococcal chemotactic factor inactivator. Infect Immun. 1985 Dec;50(3):757–764. doi: 10.1128/iai.50.3.757-764.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Wu S., de Lencastre H., Tomasz A. Sigma-B, a putative operon encoding alternate sigma factor of Staphylococcus aureus RNA polymerase: molecular cloning and DNA sequencing. J Bacteriol. 1996 Oct;178(20):6036–6042. doi: 10.1128/jb.178.20.6036-6042.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Xu J., Johnson R. C. Identification of genes negatively regulated by Fis: Fis and RpoS comodulate growth-phase-dependent gene expression in Escherichia coli. J Bacteriol. 1995 Feb;177(4):938–947. doi: 10.1128/jb.177.4.938-947.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]