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Characterization of a replication locus and formation of a higher-order complex between RepA protein and two inverted repeats in Streptomyces plasmid pSV1.

Authors
  • Chen, Yalan
  • Tan, Huarong
  • Qin, Zhongjun
Type
Published Article
Journal
FEMS Microbiology Letters
Publisher
Oxford University Press
Publication Date
Dec 01, 2013
Volume
349
Issue
2
Pages
144–152
Identifiers
DOI: 10.1111/1574-6968.12307
PMID: 24152230
Source
Medline
Keywords
License
Unknown

Abstract

We identified the minimal locus of 163-kb plasmid pSV1 of Streptomyces violaceoruber for the replication in S. lividans. This locus comprised a repA gene and an upstream 407-bp sequence containing two inverted repeats (IR-III and IR-IV) within an iteron, an AT-rich region and a 300-bp noncoding sequence (NCS). RepA protein bound specifically to a 94-bp sequence covering the intact IR-III and IR-IV to form multimers of DNA/protein complexes, but was unable to bind specifically to the NCS and the promoter of repA gene. Interestingly, this 'bound' region also leaves eight 1-bp 'unbound' spacers at 7-11-9-11-9-11-9-11-8-bp intervals. RepA protein-protein interaction could form dimers or trimers in vitro. These results suggest that a higher-order complex between pSV1 RepA protein and the long inverted repeats may be formed during the initiation of plasmid replication.

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