Volume 271, Number 36, Issue of September 6, 1996 pp. 21828-21834
©1996 by The American Society for Biochemistry and Molecular Biology, Inc.

---

Characterization of the Primary  Factor of Staphylococcus aureus

(Received for publication, May 21, 1996, and in revised form, June 27, 1996)

From the Department of Microbiology and Immunology, University of Illinois College of Medicine, Chicago, Illinois 60612

RNA polymerase (RNAP) isolated from Staphylococcus aureus is deficient in  factor and is poorly active in transcription assays. Based on amino acid sequence homology of the Bacillus subtilis vegetative  factor ^A and the predicted product of the chromosomally located plaC gene of S. aureus, it was hypothesized that plaC could encode the vegetative  factor. We cloned plaC under a T7 promoter and overexpressed it in Escherichia coli strain BL21(DE3)pLysE. The overproduced protein, present in inclusion bodies, was solubilized with guanidine hydrochloride, renatured, and purified by DEAE-Sephacel and Sephadex G-75 chromatography. The purified protein, designated ^SA, cross-reacted with the B. subtilis anti-^A antibody. E. coli core RNAP, reconstituted with ^SA, initiated promoter-specific transcription from the S. aureus promoters hla, sea, and sec and from the E. coli promoters rpoH P1, rpoH P4, and ColE1 RNA-1, which are recognized by the E. coli ⁷⁰. ^SA, when added to the purified RNAP from S. aureus, stimulated transcriptional activity of the RNAP up to 72-fold. As determined by primer extension studies, the 5-ends of the ^SA-initiated mRNAs synthesized in vitro from the agr P2 and sea promoters are in general agreement with the 5-ends of the cellular RNAs. Disruption of the plaC gene on the S. aureus chromosome was lethal. We conclude that plaC encodes the primary  factor in S. aureus.

CiteULike    Complore    Connotea    Del.icio.us    Digg    Reddit    Technorati    What's this?

This article has been cited by other articles:

				T. Koprivnjak, V. Mlakar, L. Swanson, B. Fournier, A. Peschel, and J. P. Weiss Cation-Induced Transcriptional Regulation of the dlt Operon of Staphylococcus aureus. J. Bacteriol., May 1, 2006; 188(10): 3622 - 3630. [Abstract] [Full Text] [PDF]

				E. Andre, L. Bastide, S. Michaux-Charachon, A. Gouby, P. Villain-Guillot, J. Latouche, A. Bouchet, M. Gualtieri, and J.-P. Leonetti Novel synthetic molecules targeting the bacterial RNA polymerase assembly J. Antimicrob. Chemother., February 1, 2006; 57(2): 245 - 251. [Abstract] [Full Text] [PDF]

				M. M. Senn, P. Giachino, D. Homerova, A. Steinhuber, J. Strassner, J. Kormanec, U. Fluckiger, B. Berger-Bachi, and M. Bischoff Molecular Analysis and Organization of the {sigma}B Operon in Staphylococcus aureus J. Bacteriol., December 1, 2005; 187(23): 8006 - 8019. [Abstract] [Full Text] [PDF]

				M. Mishra and R. Deora Mode of Action of the Bordetella BvgA Protein: Transcriptional Activation and Repression of the Bordetella bronchiseptica bipA Promoter J. Bacteriol., September 15, 2005; 187(18): 6290 - 6299. [Abstract] [Full Text] [PDF]

				H. Zhang, K. Morikawa, T. Ohta, and Y. Kato In vitro resistance to the CS{alpha}{beta}-type antimicrobial peptide ASABF- is conferred by overexpression of sigma factor sigB in Staphylococcus aureus J. Antimicrob. Chemother., May 1, 2005; 55(5): 686 - 691. [Abstract] [Full Text] [PDF]

				L. Shaw, E. Golonka, J. Potempa, and S. J. Foster The role and regulation of the extracellular proteases of Staphylococcus aureus Microbiology, January 1, 2004; 150(1): 217 - 228. [Abstract] [Full Text] [PDF]

				F. M. McAleese and T. J. Foster Analysis of mutations in the Staphylococcus aureus clfB promoter leading to increased expression Microbiology, January 1, 2003; 149(1): 99 - 109. [Abstract] [Full Text] [PDF]

				R. Deora Differential Regulation of the Bordetella bipA Gene: Distinct Roles for Different BvgA Binding Sites J. Bacteriol., December 15, 2002; 184(24): 6942 - 6951. [Abstract] [Full Text] [PDF]

				S. K. Chakrabarti and T. K. Misra SarA Represses agr Operon Expression in a Purified In Vitro Staphylococcus aureus Transcription System J. Bacteriol., October 15, 2000; 182(20): 5893 - 5897. [Abstract] [Full Text]

				M. S. Lee and D. A. Morrison Identification of a New Regulator in Streptococcus pneumoniae Linking Quorum Sensing to Competence for Genetic Transformation J. Bacteriol., August 15, 1999; 181(16): 5004 - 5016. [Abstract] [Full Text]

				P. F. Chan, S. J. Foster, E. Ingham, and M. O. Clements The Staphylococcus aureus Alternative Sigma Factor sigma B Controls the Environmental Stress Response but Not Starvation Survival or Pathogenicity in a Mouse Abscess Model J. Bacteriol., December 1, 1998; 180(23): 6082 - 6089. [Abstract] [Full Text]

				M. G. Kramer, M. Espinosa, T. K. Misra, and S. A. Khan Lagging strand replication of rolling-circle plasmids: Specific recognition of the ssoA-type origins in different gram-positive bacteria PNAS, September 1, 1998; 95(18): 10505 - 10510. [Abstract] [Full Text] [PDF]