Cloning and physical mapping of the Staphylococcus aureus rplL, rpoB and rpoC genes, encoding ribosomal protein L7/L12 and RNA polymerase subunits   and  '

Aboshkiwa, Mohamed A; Al‐Ani, Bahjat; Coleman, G.; Rowland, Geoffrey C.

doi:10.1099/00221287-138-9-1875

Cited by 23 publications

(19 citation statements)

References 16 publications

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“…Our Western blot results agree with data suggesting that the subunits of S. aureus RNAP have homology to the subunits of E. coli holoenzyme E 70 (2,28). The data suggest that the 165-kDa and 130-kDa proteins correspond to the ␤ and ␤Ј subunits, respectively, with sizes generally in agreement with the expected values for the gene products of the S. aureus rpoB and rpoC genes, respectively (3).…”

Section: Discussionsupporting

confidence: 82%

“…2A, lanes 2 and 3). The data are consistent with the 165-kDa and 130-kDa proteins being the gene products of the S. aureus rpoB and rpoC genes, respec- tively, as these approximate the sizes predicted from sequence analysis of these genes (142 and 129 kDa [2]). The RNAP ␣-subunit homolog is probably the 47-kDa protein, but the gene for this protein has yet to be cloned from S. aureus.…”

Section: Plasmidssupporting

confidence: 73%

“…The genes encoding the ␤ and ␤Ј subunits of S. aureus RNAP have been identified on the basis of sequence homology to the E. coli ␤ and ␤Ј genes and have predicted protein products of 142 and 129 kDa, respectively (2). The plaC gene encodes a 42-kDa protein which may be a factor of S. aureus, since it has 79% sequence identity with sigA, a major vegetative factor in Bacillus subtilis (5).…”

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In vitro transcription of pathogenesis-related genes by purified RNA polymerase from Staphylococcus aureus

1995

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The RNA polymerase (RNAP) holoenzyme of Staphylococcus aureus was purified by DNA affinity, gel filtration, and ion-exchange chromatography. This RNAP contained four major subunits with apparent molecular masses of 165, 130, 60, and 47 kDa. All four subunits of the RNAP were serologically related to the subunits of Escherichia coli E 70 holoenzyme by Western immunoblot analysis. The 60-kDa subunit was subsequently isolated and found to react with a monoclonal antibody specific to the E. coli 70 subunit. This 70 -related protein allowed E. coli core RNAP promoter-specific initiation and increased transcription by S. aureus RNAP that is unsaturated with sigma. We therefore suggest that this 60-kDa protein is a sigma factor. Purified S. aureus RNAP transcribed from the promoters of several important S. aureus virulence genes (sea, sec, hla, and agr P2) in vitro. The in vitro transcription start sites of the sea, sec, and agr P2 promoters, mapped by primer extension, were similar to those identified in vivo. The putative promoter hexamers of these three genes showed strong sequence similarity to the E. coli 70 consensus promoter, and transcription by E 70 from some of these promoters has been observed. Conversely, S. aureus RNAP does not transcribe from all E. coli 70 -dependent promoters. Taken together, our results indicate that the promoter sequences recognized by purified S. aureus RNAP are similar but not identical to those recognized by E. coli E 70 .Staphylococcus aureus is an important human and animal pathogen. It causes a variety of human infections, from localized skin suppuration to life-threatening septicemia. This pathogen also causes mastitis among dairy cattle, sheep, and goats (3, 16). In addition, staphylococcal enterotoxins are the emetic toxins responsible for staphylococcal food poisoning (6). Therefore, studying virulence gene regulation in S. aureus is useful for understanding pathogenesis and medical treatment.Many virulence genes of S. aureus have been cloned and sequenced. Known virulence factors in S. aureus include ␣-hemolysin (encoded by hla), staphylococcal enterotoxin A (sea), and staphylococcal enterotoxin C (sec). The global accessory gene regulator (agr) regulates hla transcription and translation and sec transcription but does not regulate sea expression (4,27,33,36,43). In addition to agr, the exoprotein regulator (xpr) and staphylococcal accessory regulator (sar) genes were recently identified as two distinct global elements (12,13,20,40), but the mechanism by which these global regulators affect gene expression has not been clearly elucidated. The putative promoter regions of some virulence genes have been identified (8,30,35). Recent mutational analysis of sea suggests that S. aureus has promoter structures similar to those recognized by Escherichia coli E 70 (8). However, promoter recognition by S. aureus RNA polymerase (RNAP) is not well understood yet because of the lack of an in vitro transcription system.The genes encoding S. aureus RNAP subunits have been reported. The ge...

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Section: Discussionsupporting

confidence: 82%

Section: Plasmidssupporting

confidence: 73%

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confidence: 99%

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In vitro transcription of pathogenesis-related genes by purified RNA polymerase from Staphylococcus aureus

1995

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“…1j by using oligonucleotide primers 208F and 3768R (Table 2j, which are complementary to DNA sequences that code for conserved amino acid sequences in the rpoC genes of E. coli and Staphylococcus aureus (1). Two overlapping PCR products were generated for Leuconostoc camosum ( Fig.…”

Section: Pcr Amplificationmentioning

confidence: 99%

“…The ipoB and rpoC genes are adjacent to each other on maps ( Fig. 1) in a transcriptional organization that is found in most bacteria (1,10,21). The large sizes of the p and p' subunits (151 and 156 kDa, respectively, in Escherichia coli), together with their ancient origin in evolutionary terms, indicate that these molecules could be immensely powerful molecular chronometers for phylogenetic analysis (11,16,17,19).…”

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Analysis of the ' Subunit of DNA-Dependent RNA Polymerase Does Not Support the Hypothesis Inferred from 16S rRNA Analysis that Oenococcus oeni (Formerly Leuconostoc oenos) Is a Tachytelic (Fast-Evolving) Bacterium

Morse

Collins

O’Hanlon

et al. 1996

International Journal of Systematic Bacteriology

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rRNA sequencing has shown that leuconostocs comprise three distinct phylogenetic lineages which have been designated separate genera (viz., the genera Leuconostoc sensu stricto, Oenococcus, and WeisseZZa). In addition, the 16s rRNA line formed by Oenococcus oeni (formerly Leuconostoc oenos) is exceptionally long; this fact, together with variations in the compositions of conserved positions in the 16s rRNA, has led to the hypothesis (D. Yang and C. R. Woese, Syst. Appl. Microbiol. 12:145-149, 1989) that this organism is a fast-evolving bacterium. Previous evidence that the leuconostocs should be divided into three genera and that 0. oeni is an example of tachytelic evolution has come solely from rRNA analyses. In this study we sequenced the rpoC gene encoding the p' subunit of DNA-dependent RNA polymerase of leuconostocs and performed a comparative phylogenetic analysis. The subdivision of the leuconostocs into three distinct lineages was confirmed by the rpoC gene data, but no evidence that indicated that 0. oeni is evolving at an extraordinary rate was found. If 0. oeni is truly tachytelic, then fast-evolving phenomena would be expected to occur throughout the whole genome, including this independent molecular chronometer.

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Staphylococcus

Peacock

2010

Topley &Amp; Wilson's Microbiology and Microbial Infections

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Cloning and physical mapping of the Staphylococcus aureus rplL, rpoB and rpoC genes, encoding ribosomal protein L7/L12 and RNA polymerase subunits and '

Cited by 23 publications

References 16 publications

In vitro transcription of pathogenesis-related genes by purified RNA polymerase from Staphylococcus aureus

In vitro transcription of pathogenesis-related genes by purified RNA polymerase from Staphylococcus aureus

Analysis of the ' Subunit of DNA-Dependent RNA Polymerase Does Not Support the Hypothesis Inferred from 16S rRNA Analysis that Oenococcus oeni (Formerly Leuconostoc oenos) Is a Tachytelic (Fast-Evolving) Bacterium

Staphylococcus

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