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EMBO J. 1992 Oct;11(10):3629-34.

DNA transcription and repressor binding affect deletion formation in Escherichia coli plasmids.

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  • 1Laboratoire de Génétique Microbienne, Institut National de la Recherche Agronomique, Jouy en Josas, France.

Abstract

Chimeric plasmids containing phage M13 and plasmid pBR322 sequences undergo deletions in Escherichia coli with a high frequency. In all plasmids one deletion endpoint is the M13 replication origin nick site. We examined the effects of transcription on the position of the other deletion end-point, by inserting in the plasmids an inducible promoter followed by a transcription terminator. Transcription dramatically affected deletions in an orientation-dependent way, such that greater than 95% of end-points were localized downstream from the inserted promoter when it faced the major plasmid transcripts. The end-points were not constrained to the transcribed region and were not affected by the orientation of pBR322 DNA replication. We propose that deletion events occur preferentially in a plasmid domain which is rendered positively supercoiled by convergent transcription. We also show that interaction of LacI repressor with the cognate operator generates a localized deletion hot spot. This hot spot is dependent on pBR322 replication, and therefore probably acts by arresting progression of DNA replication.

PMID:
1396563
[PubMed - indexed for MEDLINE]
PMCID:
PMC556822
Free PMC Article
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