Home LiteratureArticle Details
PMID: 15126464 Published · ppublish English Journal Article Research Support, U.S. Gov't, Non-P.H.S. Research Support, U.S. Gov't, P.H.S.

Transcription of the Staphylococcus aureus cid and lrg murein hydrolase regulators is affected by sigma factor B.

Journal of bacteriology ·Vol. 186 ·No. 10 ·2004-05-00 ·Pages 3029-37

Rice KC, Patton T, Yang SJ, Dumoulin A, Bischoff M, Bayles KW

Abstract

The Staphylococcus aureus lrg and cid loci are homologous operons that have been shown to regulate murein hydrolase activity and affect sensitivity to penicillin. Although the mode of action of these operons has not been demonstrated, a model based on the similarities of the lrgA and cidA gene products to the bacteriophage holin family of proteins has been proposed. In this study, the transcription organization and regulation of these operons were examined by Northern blot analyses. Unexpectedly, cidB and a gene located immediately downstream, designated cidC, were found to be cotranscribed on a 2.7-kb transcript. Maximal cidBC transcription occurred during early exponential growth, and high-level transcription of cidBC was dependent on the rsbU-mediated activation of the alternative sigma factor B (sigmaB). In contrast, lrgAB transcription in stationary phase was negatively regulated by sigmaB. Although cidABC transcription was not detected by Northern blot analysis, reverse transcriptase PCR revealed that these genes are also cotranscribed as a single RNA message in early exponential growth. Primer extension analysis revealed the presence of two cidBC transcription start sites, but no apparent sigmaB-dependent promoter consensus sequence was identified in these regions. The rsbU gene was also shown to have a positive impact on murein hydrolase activity but a negligible effect on sensitivity to penicillin-induced killing. These results suggest that the lrgAB and cidBC genes may be part of the S. aureus sigmaB-controlled stress regulon.

MeSH Terms
Bacterial Proteins/genetics,physiology Base Sequence Gene Expression Regulation, Bacterial Membrane Proteins/genetics Molecular Sequence Data N-Acetylmuramoyl-L-alanine Amidase/metabolism Operon Penicillins/pharmacology Promoter Regions, Genetic Sigma Factor/physiology Staphylococcus aureus/drug effects,genetics Transcription, Genetic
Chemicals
Bacterial Proteins LrgA protein, Staphylococcus aureus Membrane Proteins Penicillins SigB protein, Bacteria Sigma Factor N-Acetylmuramoyl-L-alanine Amidase
Authors & Affiliations
6 authors, click to expand affiliations / ORCID
Rice Kelly C
Department of Microbiology, Molecular Biology, and Biochemistry, University of Idaho, Moscow, Idaho 83844-3052, USA.
Patton Toni
Yang Soo-Jin
Dumoulin Alexis
Bischoff Markus
Bayles Kenneth W
References (46)
46 references, click to expand
  1. Role of the accessory gene regulator (agr) in pathogenesis of staphylococcal osteomyelitis.
    Infect Immun. 1995 Sep;63(9):3373-80 PMID: 7642265
  2. Impact of sigB mutation on Staphylococcus aureus oxacillin and vancomycin resistance varies with parental background and method of assessment.
    Int J Antimicrob Agents. 2003 Mar;21(3):256-61 PMID: 12636988
  3. Sigma(B) activity depends on RsbU in Staphylococcus aureus.
    J Bacteriol. 2001 Mar;183(6):1843-52 PMID: 11222581
  4. In vitro serial passage of Staphylococcus aureus: changes in physiology, virulence factor production, and agr nucleotide sequence.
    J Bacteriol. 2002 Mar;184(5):1430-7 PMID: 11844774
  5. Identification of LytSR-regulated genes from Staphylococcus aureus.
    J Bacteriol. 1996 Oct;178(19):5810-2 PMID: 8824633
  6. sigmaB modulates virulence determinant expression and stress resistance: characterization of a functional rsbU strain derived from Staphylococcus aureus 8325-4.
    J Bacteriol. 2002 Oct;184(19):5457-67 PMID: 12218034
  7. Analysis of genetic elements controlling Staphylococcus aureus lrgAB expression: potential role of DNA topology in SarA regulation.
    J Bacteriol. 2000 Sep;182(17):4822-8 PMID: 10940023
  8. Two bactericidal targets for penicillin in pneumococci: autolysis-dependent and autolysis-independent killing mechanisms.
    Antimicrob Agents Chemother. 1990 Jan;34(1):33-9 PMID: 1691615
  9. New members of the Escherichia coli sigmaE regulon identified by a two-plasmid system.
    FEMS Microbiol Lett. 2003 Aug 8;225(1):1-7 PMID: 12900013
  10. Optimization of a two-plasmid system for the identification of promoters recognized by RNA polymerase containing Staphylococcus aureus alternative sigma factor sigmaB.
    FEMS Microbiol Lett. 2004 Mar 19;232(2):173-9 PMID: 15033236
  11. The alternative sigma factor sigmaB in Staphylococcus aureus: regulation of the sigB operon in response to growth phase and heat shock
    Arch Microbiol. 1997 Mar 7;167(2/3):151-9 PMID: 9042755
  12. Programmed death in bacteria.
    Microbiol Mol Biol Rev. 2000 Sep;64(3):503-14 PMID: 10974124
  13. Cloning, characterization, and sequencing of an accessory gene regulator (agr) in Staphylococcus aureus.
    J Bacteriol. 1988 Sep;170(9):4365-72 PMID: 2457579
  14. Global analysis of the general stress response of Bacillus subtilis.
    J Bacteriol. 2001 Oct;183(19):5617-31 PMID: 11544224
  15. Mechanism of elementary catalytic steps of pyruvate oxidase from Lactobacillus plantarum.
    Biochemistry. 2000 Sep 5;39(35):10747-54 PMID: 10978159
  16. The Staphylococcus aureus cidAB operon: evaluation of its role in regulation of murein hydrolase activity and penicillin tolerance.
    J Bacteriol. 2003 Apr;185(8):2635-43 PMID: 12670989
  17. Strain-dependent differences in the regulatory roles of sarA and agr in Staphylococcus aureus.
    Infect Immun. 2002 Feb;70(2):470-80 PMID: 11796572
  18. Holins: the protein clocks of bacteriophage infections.
    Annu Rev Microbiol. 2000;54:799-825 PMID: 11018145
  19. Purification and biochemical characterization of pyruvate oxidase from Lactobacillus plantarum.
    J Bacteriol. 1984 Oct;160(1):273-8 PMID: 6480556
  20. Involvement of N-acetylmuramyl-L-alanine amidases in cell separation and antibiotic-induced autolysis of Escherichia coli.
    Mol Microbiol. 2001 Jul;41(1):167-78 PMID: 11454209
  21. Deletion of the alternative sigma factor sigmaB in Staphylococcus aureus reveals its function as a global regulator of virulence genes.
    J Bacteriol. 1998 Sep;180(18):4814-20 PMID: 9733682
  22. Identification and molecular characterization of a gene homologous to epr (endopeptidase resistance gene) in Staphylococcus aureus.
    Gene. 1998 Dec 11;224(1-2):67-75 PMID: 9931440
  23. The Staphylococcus aureus lrgAB operon modulates murein hydrolase activity and penicillin tolerance.
    J Bacteriol. 2000 Apr;182(7):1794-801 PMID: 10714982
  24. Teicoplanin stress-selected mutations increasing sigma(B) activity in Staphylococcus aureus.
    Antimicrob Agents Chemother. 2001 Jun;45(6):1714-20 PMID: 11353616
  25. Overexpression of sigma factor, sigma(B), urges Staphylococcus aureus to thicken the cell wall and to resist beta-lactams.
    Biochem Biophys Res Commun. 2001 Oct 26;288(2):385-9 PMID: 11606054
  26. Genetic and biochemical analyses of Escherichia coli strains having a mutation in the structural gene (poxB) for pyruvate oxidase.
    J Bacteriol. 1983 May;154(2):756-62 PMID: 6341362
  27. Regulation of sigmaB-dependent transcription of sigB and asp23 in two different Staphylococcus aureus strains.
    Mol Gen Genet. 1999 Apr;261(3):558-66 PMID: 10323238
  28. Nucleotide sequence and deduced amino acid sequence of Escherichia coli pyruvate oxidase, a lipid-activated flavoprotein.
    Nucleic Acids Res. 1986 Jul 11;14(13):5449-60 PMID: 3016647
  29. Expression of Escherichia coli pyruvate oxidase (PoxB) depends on the sigma factor encoded by the rpoS(katF) gene.
    Mol Microbiol. 1994 Mar;11(6):1019-28 PMID: 8022274
  30. A two-plasmid system for identification of promoters recognized by RNA polymerase containing extracytoplasmic stress response sigma(E) in Escherichia coli.
    J Microbiol Methods. 2001 Jun;45(2):103-11 PMID: 11311395
  31. Properties of a cryptic high-frequency transducing phage in Staphylococcus aureus.
    Virology. 1967 Sep;33(1):155-66 PMID: 4227577
  32. Description of staphylococcus serine protease (ssp) operon in Staphylococcus aureus and nonpolar inactivation of sspA-encoded serine protease.
    Infect Immun. 2001 Jan;69(1):159-69 PMID: 11119502
  33. Opposing roles of the Staphylococcus aureus virulence regulators, Agr and Sar, in Triton X-100- and penicillin-induced autolysis.
    J Bacteriol. 1998 Jul;180(14):3724-6 PMID: 9658022
  34. Death's toolbox: examining the molecular components of bacterial programmed cell death.
    Mol Microbiol. 2003 Nov;50(3):729-38 PMID: 14617136
  35. Tight regulation, modulation, and high-level expression by vectors containing the arabinose PBAD promoter.
    J Bacteriol. 1995 Jul;177(14):4121-30 PMID: 7608087
  36. Staphylococcus aureus resistant to vancomycin--United States, 2002.
    MMWR Morb Mortal Wkly Rep. 2002 Jul 5;51(26):565-7 PMID: 12139181
  37. Are the molecular strategies that control apoptosis conserved in bacteria?
    Trends Microbiol. 2003 Jul;11(7):306-11 PMID: 12875813
  38. A method to isolate RNA from gram-positive bacteria and mycobacteria.
    Anal Biochem. 1994 Nov 1;222(2):511-4 PMID: 7532381
  39. Molecular cloning, sequencing, and expression of lytM, a unique autolytic gene of Staphylococcus aureus.
    J Bacteriol. 1997 Jun;179(11):3625-31 PMID: 9171409
  40. Molecular cloning of the gene (poxB) encoding the pyruvate oxidase of Escherichia coli, a lipid-activated enzyme.
    J Bacteriol. 1984 Dec;160(3):1088-92 PMID: 6209262
  41. Influence of a functional sigB operon on the global regulators sar and agr in Staphylococcus aureus.
    J Bacteriol. 2001 Sep;183(17):5171-9 PMID: 11489871
  42. 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):6036-42 PMID: 8830703
  43. A Staphylococcus aureus autolysin that has an N-acetylmuramoyl-L-alanine amidase domain and an endo-beta-N-acetylglucosaminidase domain: cloning, sequence analysis, and characterization.
    Proc Natl Acad Sci U S A. 1995 Jan 3;92(1):285-9 PMID: 7816834
  44. Staphylococcal coagulase; mode of action and antigenicity.
    J Gen Microbiol. 1952 Feb;6(1-2):95-107 PMID: 14927856
  45. Identification of sigma(B)-dependent genes in Bacillus subtilis using a promoter consensus-directed search and oligonucleotide hybridization.
    J Bacteriol. 1999 Sep;181(18):5718-24 PMID: 10482513
  46. A new two-component regulatory system involved in adhesion, autolysis, and extracellular proteolytic activity of Staphylococcus aureus.
    J Bacteriol. 2000 Jul;182(14):3955-64 PMID: 10869073
Article Info
Journal
Journal of bacteriology
Abbr.
J Bacteriol
ISSN
0021-9193
Published
2004-05-00
Pages
3029-37
Language
English
Region
United States
NLM ID
2985120R
PMCID
PMC400629
Subset
IM
Grants
NCRR NIH HHS · P20 RR015587 · United States
NIAID NIH HHS · R01 AI038901 · United States
NCRR NIH HHS · P20RR15587 · United States
NIAID NIH HHS · R01-AI038901 · United States
Analysis Services
Analysis Services

Contact

No. 2 Wenbo Road, Zhangqiu District, Jinan, Shandong

Qilu Normal University · Genelibs Bioinformatics Lab

750 Shunhua Rd, Jinan

2F, Bldg F, University Science Park

Tel: 0531-88819269

WeChat Official Account

Follow our WeChat subscription account for real-time updates and the latest in medical and biological research.


Business Email

E-mail: product@genelibs.com