Fractionation of 32P-labelled ribonucleic acids on polyacrylamide gels and their characterization by fingerprinting

Hindley, J. Roger

doi:10.1016/0022-2836(67)90248-3

Cited by 146 publications

(68 citation statements)

References 10 publications

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“…Evidence of the existence of sRNAs other than 5S rRNA and tRNAs came with the development of PAGE to analyze total RNA (11). Besides tRNAs, 5S, 16S, and 23S rRNAs, additional bands can be detected by ethidium bromide staining (Fig.…”

Section: Resultsmentioning

confidence: 99%

Small RNA genes expressed from Staphylococcus aureus genomic and pathogenicity islands with specific expression among pathogenic strains

Pichon

Felden

2005

Proc. Natl. Acad. Sci. U.S.A.

234

271

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Small RNA (sRNA) genes are expressed in all organisms, primarily as regulators of translation and message stability. We have developed comparative genomic approaches to identify sRNAs that are expressed by Staphylococcus aureus , the most common cause of hospital-acquired infections. This study represents an in-depth analysis of the RNome of a Gram-positive bacterium. A set of sRNAs candidates were identified in silico within intergenic regions, and their expression levels were monitored by using microarrays and confirmed by Northern blot hybridizations. Two sRNAs were also detected directly from purification and RNA sequence determination. In total, at least 12 sRNAs are expressed from the S. aureus genome, five from the core genome and seven from pathogenicity islands that confer virulence and antibiotic resistance. Three sRNAs are present in multiple (two to five) copies. For the sRNAs that are conserved throughout the bacterial phylogeny, their secondary structures were inferred by phylogenetic comparative methods. In vitro binding assays indicate that one sRNA encoded within a pathogenicity island is a trans-encoded antisense RNA regulating the expression of target genes at the posttranscriptional level. Some of these RNAs show large variations of expression among pathogenic strains, suggesting that they are involved in the regulation of staphylococcal virulence.

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Section: Resultsmentioning

confidence: 99%

Small RNA genes expressed from Staphylococcus aureus genomic and pathogenicity islands with specific expression among pathogenic strains

Pichon

Felden

2005

Proc. Natl. Acad. Sci. U.S.A.

234

271

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“…1-3). 6S RNA was one of the first bacterial sRNAs to be discovered 4,5 and is one of the most abundant cellular RNAs. Yet strikingly, it has escaped functional analysis for decades, and was only lately discovered to be ubiquitous in bacteria.…”

Section: Introductionmentioning

confidence: 99%

In vivo and in vitro analysis of 6S RNA-templated short transcripts inBacillus subtilis

Beckmann

Burenina²,

Hoch³

et al. 2011

RNA Biology

123

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“…Escherichia coli 6S RNA was identified in the late 1960's as a highly abundant species in stationary phase cells and was one of the first RNAs to be sequenced (Hindley 1967;Brownlee 1971). This RNA is now known to regulate hundreds of s 70 (housekeeping)-dependent promoters in E. coli (Wassarman and Storz 2000;Trotochaud and Wassarman 2005;Cavanagh et al 2008).…”

Section: Introductionmentioning

confidence: 99%

Binding and release of the 6S transcriptional control RNA

Shephard¹,

Dobson²,

Unrau³

2010

RNA

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ABSTRACT6S RNA is an important noncoding RNA that regulates eubacterial transcription. In Escherichia coli this RNA binds to the s 70 RNA polymerase holoenzyme and is released by the synthesis of a short product RNA. In order to determine how binding and release are controlled by the 6S RNA sequence, we used in vitro selection to screen a high diversity library containing ;4 3 10 12 sequences for functional 6S RNA variants. Residues critical for binding were found to be located in a ''-35'' region upstream of the 6S RNA transcription bubble mimic structure. Mutating these phylogenetically conserved residues invariably led to decreases in binding and removing them abolished binding, implicating these nucleotides in a biologically important interaction with the Es 70 complex. Interestingly, mutation of phylogenetically conserved ''-10'' residues that were also upstream of the site of pRNA synthesis was found to influence 6S RNA release rates in addition to modulating -35 binding. These results indicate how 6S RNA -35 binding to s 70 RNA polymerase holoenzyme can regulate expression from ''strong'' and ''weak'' -35 DNA promoters and suggest that 6S RNA release rates have been fine tuned over evolutionary time so as to correctly regulate cellular levels of transcription.

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Fractionation of 32P-labelled ribonucleic acids on polyacrylamide gels and their characterization by fingerprinting

Cited by 146 publications

References 10 publications

Small RNA genes expressed from Staphylococcus aureus genomic and pathogenicity islands with specific expression among pathogenic strains

Small RNA genes expressed from Staphylococcus aureus genomic and pathogenicity islands with specific expression among pathogenic strains

In vivo and in vitro analysis of 6S RNA-templated short transcripts inBacillus subtilis

Binding and release of the 6S transcriptional control RNA

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