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Protein RepC is involved in copy number control of the broad host range plasmid RSF1010

Haring, V and Scholz, P and Scherzinger, E and Frey, J and Derbyshire, K and Hatfull, G and Willetts, NS and Bagdasarian, M (1985) Protein RepC is involved in copy number control of the broad host range plasmid RSF1010. Proceedings of the National Academy of Sciences of the United States of America, 82 (18). 6090 - 6094. ISSN 0027-8424

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Abstract

Essential replication (rep) genes of the broad host range plasmid RSF1010 have been cloned onto controlled expression vectors and their protein products have been visualized, after induction, by NaDodSO4/polyacrylamide gel electrophoresis of whole cell lysates. During this induction the replication of a coresident RSF1010 replicon, pKT210, was analyzed by quantitative DNA-DNA hybridization. The initiation of pKT210 replication was stimulated 6-fold by a simultaneous overproduction of the RepA and RepC proteins compared to cells in which only the RepA protein was overproduced. An enhanced synthesis of the RepB protein resulted in a 1.6-fold stimulation of pKT210 replication, whereas an overproduction of the RepA protein alone had no effect. Purified RepC protein has been shown to bind preferentially to DNA carrying the replication origin of RSF1010. Within this segment it was bound specifically to those DNA fragments that contained the 20-base-pair repeats of the origin region. These results suggest that RepC protein acts as a positive replication regulator, that its concentration is rate limiting, and that the replication rate of RSF1010 is controlled, at least in part, at the level of RepC synthesis.


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Details

Item Type: Article
Status: Published
Creators/Authors:
CreatorsEmailPitt UsernameORCID
Haring, V
Scholz, P
Scherzinger, E
Frey, J
Derbyshire, K
Hatfull, Ggfh@pitt.eduGFH
Willetts, NS
Bagdasarian, M
Date: 1 December 1985
Date Type: Publication
Journal or Publication Title: Proceedings of the National Academy of Sciences of the United States of America
Volume: 82
Number: 18
Page Range: 6090 - 6094
DOI or Unique Handle: 10.1073/pnas.82.18.6090
Schools and Programs: Dietrich School of Arts and Sciences > Biological Sciences
Refereed: Yes
ISSN: 0027-8424
MeSH Headings: Bacterial Proteins--genetics; Base Sequence; Cloning, Molecular; DNA Replication; DNA, Bacterial--genetics; DNA-Binding Proteins--genetics; Escherichia coli--genetics; Gene Expression Regulation; Plasmids
Other ID: NLM PMC390705
PubMed Central ID: PMC390705
PubMed ID: 3898079
Date Deposited: 30 Oct 2015 15:05
Last Modified: 02 Feb 2019 15:57
URI: http://d-scholarship.pitt.edu/id/eprint/26245

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