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PMID: 2193921 Published · ppublish English Journal Article Research Support, U.S. Gov't, P.H.S.

Mutations in the trfA replication gene of the broad-host-range plasmid RK2 result in elevated plasmid copy numbers.

Journal of bacteriology ·Vol. 172 ·No. 7 ·1990-07-00 ·Pages 3859-67

Durland RH, Toukdarian A, Fang F, Helinski DR

Abstract

Mutated forms of trfA, the replication protein gene of plasmid RK2, that support a minimal RK2 origin plasmid in Escherichia coli at copy numbers up to 23-fold higher than normal have been isolated. Six such high-copy-number (copy-up) mutations were mapped and sequenced. In each case, a single base transition led to an amino acid substitution in the TrfA protein primary sequence. The six mutations affected different residues of the protein and were located within a 69-base-pair region encoding 24 amino acids. Dominance tests showed that each of the mutants can be suppressed by wild-type trfA in trans, but suppression is highly dependent on the amount of wild-type protein produced. Excess mutant TrfA protein provided in trans significantly increased the copy number of RK2 and other self-replicating derivatives of RK2 that contain a wild-type trfA gene. These observations suggest that the mutations affect a regulatory activity of the TrfA replication protein that is a key factor in the control of initiation of RK2 replication.

MeSH Terms
Alleles Amino Acid Sequence Bacterial Proteins/genetics Base Sequence Escherichia coli/genetics Genes, Bacterial Molecular Sequence Data Mutation Plasmids/genetics Restriction Mapping
Chemicals
Bacterial Proteins
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Durland R H
Center for Molecular Genetics, University of California, San Diego, La Jolla 92093.
Toukdarian A
Fang F
Helinski D R
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Article Info
Journal
Journal of bacteriology
Abbr.
J Bacteriol
ISSN
0021-9193
DOI
10.1128/jb.172.7.3859-3867.1990
Published
1990-07-00
Pages
3859-67
Language
English
Region
United States
NLM ID
2985120R
PMCID
PMC213367
Subset
IM
Grants
NIAID NIH HHS · AI-07194 · United States
External Links
PubMed source DOI: 10.1128/jb.172.7.3859-3867.1990 PMC Full Text View journal details
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