Skip to main content
Log in

Plasmid multimerization is dependent on RAD52 activity in Saccharomyces cerevisiae

  • Short Communications
  • Published:
Molecular and General Genetics MGG Aims and scope Submit manuscript

Summary

A mutant plasmid, pX, derived from the 1453 base pair small plasmid, YARp1 (or TRP1 RI circle), consists of 849 base pairs of DNA bearing the TRP1 gene and the ARS1 sequence of Saccharomyces cerevisiae and, unlike YARp1 and other commonly used yeast plasmids, highly multimerizes in a S. cerevisiae host. The multimerization of pX was dependent on RAD52, which is known to be necessary for homologous recombination in S. cerevisiae. Based upon this observation, a regulated system of multimerization of pX with GAL1 promoter-driven RAD52 has been developed. We conclude that the regulated multimerization of pX could provide a useful model system to study genetic recombination in the eukaryotic cell, in particular to investigate recombination intermediates and the effects of various trans-acting mutations on the multimerization and recombination of plasmids.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

References

  • Adzuma K, Ogawa T, Ogawa H (1984) Primary structure of the RAD52 gene in Saccharomyces cerevisiae. Mol Cell Biol 4:2735–2744

    Google Scholar 

  • Bolivar F, Rodriguez RL, Betlach MC, Boyer HW (1977) Construction and characterization of new cloning vehicles. I. Ampicillin-resistant derivatives of the plasmid pMB9. Gene 2:75–93

    Google Scholar 

  • Broach JR, Strathern JN, Hicks JB (1979) Transformation in yeast: development of a hybrid cloning vector and isolation of the CAN1 gene. Gene 8:121–133

    Google Scholar 

  • Broach JR, Li Y-Y, Wu L-CC, Jayaram MM (1983) Vectors for high-level inducible expression of cloned genes in yeast. In: Inouye M (ed) Experimental manipulation of gene expression. Academic Press, New York

    Google Scholar 

  • Cohen A, Laban A (1983) Plasmidic recombination in Escherichia coli K12. Mol Gen Genet 189:471–474

    Google Scholar 

  • Embretson JE, Livingston DM (1984) A plasmid model to study genetic recombination in yeast. Gene 29:293–302

    Google Scholar 

  • Fishel RA, James AA, Kolodner R (1981) recA independent general recombination of plasmids. Nature 294:184–186

    Google Scholar 

  • Game JC, Zamb TJ, Braun RJ, Resnick M, Roth RM (1980) The role of radiation (rad) genes of meiotic recombination in yeast. Genetics 94:51–68

    Google Scholar 

  • Hakkaart MJ, van der Elzen PJM, Veltkamp E, Nijkamp HJJ (1984) Maintenance of multicopy plasmid Clo DF13 in E. coli cells: evidence for site-specific recombination at parB. Cell 36:203–209

    Google Scholar 

  • Harashima S, Takagi A, Oshima Y (1984) Transformation of protoplasted yeast cells is directly associated with cell fusion. Mol Cell Biol 4:771–778

    Google Scholar 

  • Harashima S, Shimada Y, Oshima Y (1985) A mutant plasmid with increased stability of holding and polymerization in Saccharomyces cerevisiae. Mol Gen Genet 199:14–20

    Google Scholar 

  • Hereford L, Fahrner K, Woolford J Jr, Rosbash M, Kaback DB (1979) Isolation of yeast histone genes H2A and H2B. Cell 18:1261–1271

    Google Scholar 

  • Ito H, Fukuda Y, Murata K, Kimura A (1983) Transformation of intact yeast cells treated with alkali cations. J Bacteriol 153:163–168

    Google Scholar 

  • James AA, Morrison PT, Kolodner R (1982) Genetic recombination of bacterial plasmid DNA. Analysis of the effect of recombination-deficient mutations on plasmid recombination. J Mol Biol 160:411–430

    Google Scholar 

  • Johnston M, Davis RW (1984) Sequences that regulate the divergent GALL-GAL10 promoter in Saccharomyces cerevisiae. Mol Cell Biol 4:1440–1448

    Google Scholar 

  • Kolodner R (1980) Genetic recombination of bacterial plasmid DNA: electron microscopic analysis of intramolecular recombination catalyzed in vitro. Proc Natl Acad Sci USA 77:4847–4851

    Google Scholar 

  • Kolodner R, Fishel RA, Howard M (1985) Genetic recombination of bacterial plasmid DNA: effect of RecF pathway mutations on plasmid recombination in Escherichia coli. J Bacteriol 163:1060–1066

    Google Scholar 

  • Laban A, Cohen A (1981) Interplasmidic and intraplasmidic recombination in Escherichia coli K12. Mol Gen Genet 184:200–207

    Google Scholar 

  • Long CH, Brajkovich CM, Scott JF (1985) Alternative model for chromatin organization of the Saccharomyces cerevisiae chromosomal DNA plasmid TRP1 RI circle (YARpl). Mol Cell Biol 5:3124–3130

    Google Scholar 

  • Maniatis T, Fritsch EF, Sambrook J (1982) Molecular cloning: a laboratory manual. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York

    Google Scholar 

  • Orr-Weaver TL, Szostak JW, Rothstein RJ (1981) Yeast transformation: a model system for the study recombination. Proc Natl Acad Sci USA 78:6354–6358

    Google Scholar 

  • Potter H, Dressler D (1977) On the mechanism of genetic recombination: the maturation of recombination intermediates. Proc Natl Acad Sci USA 74:4168–4172

    Google Scholar 

  • Prakash S, Prakash L, Burke W, Montelone BA (1980) Effects of the RAD52 gene on recombination in Saccharomyces cerevisiae. Genetics 94:31–50

    Google Scholar 

  • Sherman F, Fink GR, Lawrence CW (1974) Methods in yeast genetics. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York

    Google Scholar 

  • Shild D, Konforti B, Perez C, Gish W, Mortimer RK (1983) Isolation and characterization of yeast DNA repair genes. I. Cloning of the RAD52 gene. Curr Genet 7:85–92

    Google Scholar 

  • Southern EM (1975) Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol 98:503–517

    Google Scholar 

  • Stinchcomb DT, Struhl K, Davis RW (1979) Isolation and characterization of a yeast chromosomal replicator. Nature 282:39–43

    Google Scholar 

  • Summers DK, Sherratt DJ (1984) Multimerization of high copy number plasmids causes instability: Co1E1 encodes a determinant essential for plasmid monomerization and stability. Cell 36:1097–1103

    Google Scholar 

  • Toh-e A, Tada S, Oshima Y (1982) 2-μm DNA-like plasmids in the osmophilic haploid yeast Saccharomyces rouxii. J Bacteriol 151:1380–1390

    Google Scholar 

  • Tschumper G, Carbon J (1980) Sequence of a yeast DNA fragment containing a chromosomal replicator and the TRP1 gene. Gene 10:157–166

    Google Scholar 

  • Whiteway MS, Ahmed A (1984) Recombinational instability of a chimeric plasmid in Saccharomyces cerevisiae. Mol Cell Biol 4:195–198

    Google Scholar 

  • Zakian VA, Scott JF (1982) Construction, replication, and chromatin structure of TRP1 RI Circle, a multiple-copy synthetic plasmid derived from Saccharomyces cerevisiae chromosomal DNA. Mol Cell Biol 2:221–232

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Additional information

Communicated by C.P. Hollenberg

Rights and permissions

Reprints and permissions

About this article

Cite this article

Harashima, S., Shimada, Y., Nakade, S. et al. Plasmid multimerization is dependent on RAD52 activity in Saccharomyces cerevisiae . Mol Gen Genet 219, 495–498 (1989). https://doi.org/10.1007/BF00259627

Download citation

  • Received:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00259627

Key words

Navigation