Plasmid pGA1 from Corynebacterium glutamicum codes for a gene product that positively influences plasmid copy number
Language English Country United States Media print
Document type Journal Article, Research Support, Non-U.S. Gov't
PubMed
9045809
PubMed Central
PMC178862
DOI
10.1128/jb.179.5.1525-1532.1997
Knihovny.cz E-resources
- MeSH
- Bacterial Proteins chemistry genetics physiology MeSH
- Corynebacterium genetics MeSH
- DNA-Binding Proteins * MeSH
- DNA Helicases chemistry genetics MeSH
- Molecular Sequence Data MeSH
- Open Reading Frames MeSH
- Plasmids genetics MeSH
- Promoter Regions, Genetic MeSH
- Gene Expression Regulation, Bacterial MeSH
- Replicon genetics MeSH
- Amino Acid Sequence MeSH
- Base Sequence MeSH
- Sequence Alignment MeSH
- Trans-Activators chemistry genetics MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
- Names of Substances
- Bacterial Proteins MeSH
- DNA-Binding Proteins * MeSH
- DNA Helicases MeSH
- ORFA protein, Corynebacterium MeSH Browser
- replication initiator protein MeSH Browser
- Trans-Activators MeSH
The complete nucleotide sequence (4,826 bp) of the cryptic plasmid pGA1 from Corynebacterium glutamicum was determined. DNA sequence analysis revealed four putative coding regions (open reading frame A [ORFA], ORFA2, ORFB, and ORFC). ORFC was identified as a rep gene coding for an initiator of plasmid replication (Rep) according to the high level of homology of its deduced amino acid sequence with the Rep proteins of plasmids pSR1 (from C. glutamicum) and pNG2 (from Corynebacterium diphtheriae). This function was confirmed by deletion mapping of the minimal replicon of pGA1 (1.7 kb) which contains only ORFC. Deletion derivatives of pGA1 devoid of ORFA exhibited significant decreases in the copy number in C. glutamicum cells and displayed segregational instability. Introduction of ORFA in trans into the cells harboring these deletion plasmids dramatically increased their copy number and segregational stability. The ORFA gene product thus positively influences plasmid copy number. This is the first report on such activity associated with a nonintegrating bacterial plasmid. The related plasmids pGA1, pSR1, and pNG2 lacking significant homology with any other plasmid seem to be representatives of a new group of plasmids replicating in the rolling-circle mode.
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General and molecular microbiology and microbial genetics in the IM CAS
Control of rep gene expression in plasmid pGA1 from Corynebacterium glutamicum
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