The activity of a single-stranded promoter of plasmid ColIb-P9 depends on its secondary structure

Molecular Microbiology
M T NasimW J Brammar

Abstract

The leading region of the conjugal bacterial plasmid ColIb-P9 contains three dispersed repeats of a 328 bp sequence homologous to Frpo, a sequence from plasmid F that acts as a promoter in single-stranded DNA. One of these sequences, ssi3, inactive in the double-stranded form, promoted in vitro transcription exclusively from the single strand that is transferred during conjugation. Promoter activity was dependent on the presence of RNA polymerase holoenzyme containing sigma 70. Transcription initiated from the position predicted from folding the single-stranded DNA to form a pseudo double-stranded hairpin structure containing recognizable -35 and -10 promoter elements. Footprinting of RNA polymerase holoenzyme on single-stranded ssi3 DNA was consistent with this suggestion. Mutagenesis of the putative -35 region inactivated the promoter, but random mutations in the -10 region had little effect. The putative -10 region is a poor match to the consensus sequence and contains mismatched bases. Elimination of these mismatches invariably destroyed single-strand promoter activity. These observations reveal the crucial contribution of the unpaired bases in the -10 region in potentiating the formation of the productive open complex with...Continue Reading

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Citations

Sep 27, 2013·Nucleic Acids Research·Wil A M LoenenGeoffrey G Wilson
Dec 2, 2010·Microbiology and Molecular Biology Reviews : MMBR·David BikardDidier Mazel
Apr 18, 2007·Proceedings of the National Academy of Sciences of the United States of America·Elena K DavydovaLucia B Rothman-Denes
Mar 23, 2013·The Biochemical Journal·Danil PupovAndrey Kulbachinskiy
Apr 1, 2017·Plasmid·Katherine E L Cox, Joel F Schildbach
Aug 2, 2005·Journal of Molecular Biology·Stephen G LandtAlan D Frankel
Dec 12, 2018·FEBS Letters·Daria EsyuninaAndrey Kulbachinskiy
Nov 26, 2008·Journal of Bacteriology·Fabián Lorenzo-Díaz, Manuel Espinosa
Nov 1, 2018·Nucleic Acids Research·Jorge Val-CalvoWilfried J J Meijer
Jul 7, 2020·Frontiers in Microbiology·Masatoshi MiyakoshiMasataka Tsuda
Dec 4, 2020·Nucleic Acids Research·Alexey FomenkovRichard J Roberts

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