Abstract
A circular DNA plasmid, pSR1, isolated from Zygosaccharomyces rouxii has a pair of inverted repeats consisting of completely homologous 959-base pair (bp) sequences. Intramolecular recombination occurs frequently at the inverted repeats in cells of Saccharomyces cerevisiae, as well as in Z. rouxii, and is catalyzed by a protein encoded by the R gene of its own genome. The recombination is, however, independent of the RAD52 gene of the host genome. A site for initiation of the intramolecular recombination in the S. cerevisiae host was delimited into, at most, a 58-bp region in the inverted repeats by using mutant plasmids created by linker insertion. The 58-bp region contains a pair with 14-bp dyad symmetry separated by a 3-bp spacer sequence. The recombination initiated at this site was accompanied by a high frequency of gene conversion (3 to 50% of the plasmid clones examined). Heterogeneity created by the linker insertion or by a deletion (at most 153 bp so far tested) at any place on the inverted repeats was converted to a homologous combination by the gene conversion, even in the rad52-1 mutant host. A mechanism implying branch migration coupled with DNA replication is discussed.
MeSH Terms
Base Sequence
Cloning, Molecular
Escherichia coli/genetics
Gene Conversion
Genes, Fungal
Molecular Sequence Data
Mutation
Plasmids
Recombination, Genetic
Saccharomyces/genetics
Saccharomyces cerevisiae/genetics
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Matsuzaki H
Department of Fermentation Technology, Faculty of Engineering, Osaka University, Japan.
Araki H
Oshima Y
References (28)
28 references, click to expand
-
Gene conversion between duplicated genetic elements in yeast.
Nature. 1981 Jul 23;292(5821):306-11
PMID: 6265790
-
Bacteriophage P1 site-specific recombination. III. Strand exchange during recombination at lox sites.
J Mol Biol. 1981 Aug 25;150(4):603-8
PMID: 6460113
-
Homology-dependent interactions in phage lambda site-specific recombination.
Nature. 1987 Sep 24-30;329(6137):346-8
PMID: 2957599
-
Recombination within the yeast plasmid 2mu circle is site-specific.
Cell. 1982 May;29(1):227-34
PMID: 6286142
-
Site-specific genetic recombination promoted by the FLP protein of the yeast 2-micron plasmid in vitro.
Cold Spring Harb Symp Quant Biol. 1984;49:797-804
PMID: 6597765
-
Physical and functional structure of a yeast plasmid, pSB3, isolated from Zygosaccharomyces bisporus.
Nucleic Acids Res. 1985 Jun 25;13(12):4267-83
PMID: 2989791
-
The FLP recombinase of the Saccharomyces cerevisiae 2 microns plasmid attaches covalently to DNA via a phosphotyrosyl linkage.
Mol Cell Biol. 1985 Nov;5(11):3274-9
PMID: 2427927
-
A system for shotgun DNA sequencing.
Nucleic Acids Res. 1981 Jan 24;9(2):309-21
PMID: 6259625
-
Site-specific recombination of yeast 2-micron DNA in vitro.
Proc Natl Acad Sci U S A. 1983 Dec;80(23):7284-8
PMID: 6316354
-
Mating type-like conversion promoted by the 2 micrograms circle site-specific recombinase: implications for the double-strand-gap repair model.
Mol Cell Biol. 1986 Nov;6(11):3831-7
PMID: 3025614
-
The FLP recombinase of the 2 micron circle DNA of yeast: interaction with its target sequences.
Cell. 1985 Apr;40(4):795-803
PMID: 3879971
-
New M13 vectors for cloning.
Methods Enzymol. 1983;101:20-78
PMID: 6310323
-
The FLP protein of the 2-micron plasmid of yeast. Purification of the protein from Escherichia coli cells expressing the cloned FLP gene.
J Biol Chem. 1985 Oct 5;260(22):12313-9
PMID: 2995370
-
Nucleotide sequence of the yeast plasmid.
Nature. 1980 Aug 28;286(5776):860-5
PMID: 6251374
-
DNA sequencing with chain-terminating inhibitors.
Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463-7
PMID: 271968
-
Identification of the crossover site during FLP-mediated recombination in the Saccharomyces cerevisiae plasmid 2 microns circle.
Mol Cell Biol. 1986 Oct;6(10):3357-67
PMID: 3540590
-
The RAD52 gene is required for homothallic interconversion of mating types and spontaneous mitotic recombination in yeast.
Proc Natl Acad Sci U S A. 1980 Jan;77(1):503-7
PMID: 6987653
-
Copy number amplification of the 2 micron circle plasmid of Saccharomyces cerevisiae.
J Theor Biol. 1986 Mar 21;119(2):197-204
PMID: 3525993
-
Site-specific recombination intermediates trapped with suicide substrates.
Cell. 1987 Aug 28;50(5):779-88
PMID: 3040260
-
Effects of the RAD52 Gene on Recombination in SACCHAROMYCES CEREVISIAE.
Genetics. 1980 Jan;94(1):31-50
PMID: 17248995
-
Construction and characterization of new cloning vehicles. II. A multipurpose cloning system.
Gene. 1977;2(2):95-113
PMID: 344137
-
Molecular and functional organization of yeast plasmid pSR1.
J Mol Biol. 1985 Mar 20;182(2):191-203
PMID: 3889347
-
The double-strand-break repair model for recombination.
Cell. 1983 May;33(1):25-35
PMID: 6380756
-
Association of reciprocal exchange with gene conversion between the repeated segments of 2-micron circle.
J Mol Biol. 1986 Oct 5;191(3):341-54
PMID: 3029381
-
In vitro systems for genetic recombination of the DNAs of bacteriophage T7 and yeast 2-micron circle.
Cold Spring Harb Symp Quant Biol. 1984;49:789-96
PMID: 6597764
-
The FLP recombinase of the yeast 2-micron plasmid: characterization of its recombination site.
Proc Natl Acad Sci U S A. 1985 Nov;82(21):7270-4
PMID: 2997780
-
2-micrometers DNA-like plasmids in the osmophilic haploid yeast Saccharomyces rouxii.
J Bacteriol. 1982 Sep;151(3):1380-90
PMID: 6286598
-
Transformation of intact yeast cells treated with alkali cations.
J Bacteriol. 1983 Jan;153(1):163-8
PMID: 6336730