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A Conserved Pattern of Primer-Dependent Transcription Initiation in and Revealed by 5′ RNA-seq


Primer-dependent transcription initiation, PDI, refers to an alternative mechanism of transcription initiation whereby the first phosphodiester bond within the nascent RNA is formed between a 2- to ~4-nt RNA primer and an incoming nucleoside triphosphate. Although PDI has been shown to occur in E. coli, the impact of PDI on E. coli physiology, and the extent to which PDI occurs in other bacteria is unknown. Here we establish that PDI modulates the ability of E. coli to form biofilms, a surface attached community of bacteria encased in a polymeric matrix. We further describe a significantly improved RNA-seq based method for the detection of PDI in cells. Using this method we document the occurrence of PDI in the pathogenic bacterium Vibrio cholerae. We further show that the pattern of PDI in V. cholerae is identical to that observed in E. coli, suggesting that PDI in these two organisms may occur through a conserved process that produces identical populations of 2- to ~4-nt RNA primers. Our findings suggest PDI may be widespread in Gammaproteobacteria.


Vyšlo v časopise: A Conserved Pattern of Primer-Dependent Transcription Initiation in and Revealed by 5′ RNA-seq. PLoS Genet 11(7): e32767. doi:10.1371/journal.pgen.1005348
Kategorie: Research Article
prolekare.web.journal.doi_sk: https://doi.org/10.1371/journal.pgen.1005348

Souhrn

Primer-dependent transcription initiation, PDI, refers to an alternative mechanism of transcription initiation whereby the first phosphodiester bond within the nascent RNA is formed between a 2- to ~4-nt RNA primer and an incoming nucleoside triphosphate. Although PDI has been shown to occur in E. coli, the impact of PDI on E. coli physiology, and the extent to which PDI occurs in other bacteria is unknown. Here we establish that PDI modulates the ability of E. coli to form biofilms, a surface attached community of bacteria encased in a polymeric matrix. We further describe a significantly improved RNA-seq based method for the detection of PDI in cells. Using this method we document the occurrence of PDI in the pathogenic bacterium Vibrio cholerae. We further show that the pattern of PDI in V. cholerae is identical to that observed in E. coli, suggesting that PDI in these two organisms may occur through a conserved process that produces identical populations of 2- to ~4-nt RNA primers. Our findings suggest PDI may be widespread in Gammaproteobacteria.


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