RNase H overproduction corrects a defect at the level of transcription elongation during rRNA synthesis in the absence of DNA topoisomerase I in Escherichia coli.
It has been suggested that the major function of DNA topoisomerase I in Escherichia coli is to suppress the formation of R-loops, which could inhibit growth. Although the currently available data suggest that the inhibitory effect of R-loops is exerted at the level of gene expression, this has never been demonstrated. In the present report, we show that rRNA synthesis is significantly impaired at the level of transcription elongation in a bacterial strain lacking DNA topoisomerase I. We found that this inhibition is due to transcriptional blocks. RNase H overproduction is also shown to considerably reduce the extent of such transcriptional blocks during rRNA synthesis. Moreover, one of these transcriptional blockage sites is located within a region where extensive R-loop formation was previously shown to occur on a plasmid DNA in the absence of DNA topoisomerase I. Together, these results allow us to propose that an important function of DNA topoisomerase I is to inhibit the formation of R-loops, which may otherwise translate into roadblocks for RNA polymerases. Our results also highlight the potential regulatory role of DNA supercoiling at the level of transcription elongation.
Hraiky
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C
Département de Microbiologie et Immunologie, Université de Montréal, Québec H3C 3J7, Canada.
Raymond
M A
MA
Drolet
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M
D004264:DNA Topoisomerases, Type I / Q000502:physiology*; D004278:DNA, Superhelical / Q000502:physiology; D004926:Escherichia coli / Q000235:genetics*; D012335:RNA, Ribosomal / Q000096:biosynthesis*; D012336:RNA, Ribosomal, 16S / Q000032:analysis; D012338:RNA, Ribosomal, 23S / Q000032:analysis; D016914:Ribonuclease H / Q000502:physiology*; D014158:Transcription, Genetic
D004278:DNA, Superhelical; D012335:RNA, Ribosomal; D012336:RNA, Ribosomal, 16S; D012338:RNA, Ribosomal, 23S; D016914:Ribonuclease H; D004264:DNA Topoisomerases, Type I