Summary
Stable maintenance of the yeast 2 μm circle requires an origin of replication and three additional plasmidencoded functions. These are the two trans-acting products of plasmid genes B and C, and the cis-acting region designated STB or REP3. We have carried out detailed studies of the mitotic stability and copy number of 2 μm-based plasmids in [cir+] and [cir0] hosts. The results obtained demonstrate that all three of these functions play a major role in plasmid partitioning. The behaviour of hybrid plasmids carrying various regions of the 2 μm circle has also been studied in [cir0] isolates containing chromosomally integrated copies of the B and/or C genes. The results demonstrate that the efficiency of partitioning is dependent upon the dosage of gene B but is independent of the dosage of gene C.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Araki H, Jearnpipatkul A, Tatsumi H, Sakurai T, Ushio K, Muta T, Oshima Y (1985) Molecular and functional organisation of the yeast plasmid pSR1. J Mol Biol 182:191–203
Austin S, Abeles A (1983a) Partition of unit-copy miniplasmids to daughter cells I. J Mol Biol 169:353–372
Austin S, Abeles A (1983b) Partition of unit-copy miniplasmids to daughter cells II. J Mol Biol 169:373–387
Beggs JD (1978) Transformation of yeast by a replicating hybrid plasmid. Nature 275:104–109
Beggs JD, Guerineau M, Atkins JF (1976) A map of the restriction targets in yeast 2-micron plasmid DNA cloned on bacteriophage Lambda. Mol Gen Genet 148:281–294
Birnboim AC, Doly J (1979) A rapid alkaline extraction procedure for screening recombinant plasmid DNA. Nucl Acids Res 76:1513–1523
Botstein D, Davis RW (1982) Principles and practice of recombinant DNA research with yeast. In: Strathern JN, Jones EW, Broach JR (eds) The molecular biology of the yeast Saccharomyces: metabolism and expression. Cold Spring Harbor Laboratory, New York, pp 607–636
Broach JR, Hicks JB (1980) Replication and recombination functions associated with the yeast plasmid 2 μ circle. Cell 21:501–508
Broach JR, Strathern JM, Hicks JB (1979) Transformation in yeast: Development of a hybrid cloning vector and isolation of the CAN1 gene. Gene 8:121–133
Clark-Walker GD, Miklos GL (1974) Localization and quantification of circular DNA in yeast. Eur J Biochem 41:359–365
Dobson MJ, Futcher AB, Cox BS (1980a) Control of recombination within and between DNA plasmids of Saccharomyces cerevisiae. Curr Genet 2:193–200
Dobson MJ, Futcher AB, Cox BS (1980b) Loss of 2 μm DNA from S. cerevisiae transformed with the chimaeric plasmid pJDB219. Current Genet 2:201–205
Dobson MJ, Kingsman SM, Kingsman AJ (1981) Sequence variation in the LEU2 region of Saccharomyces cerevisiae genome. Gene 16:133–139
Feinberg AP, Vogelstein B (1983) A technique for radiolabelling DNA restriction endonuclease fragments to high specific activity. Anal Biochem 132:6–13
Hartley JL, Donelson JE (1980) Nucleotide sequence of the yeast plasmid. Nature 286:860–864
Hollenberg CP, Borst P, Van Bruggen EFJ (1970) Mitochondrial DNA V. A 25 μ closed circular duplex DNA molecule in wildtype yeast mitochondria. Structure and genetic complexity. Biochim Biophys Acta 209:1–15
Hsiao C-L, Carbon J (1981) Direct selection procedure for the isolation of functional centromeric DNA. Proc Natl Acad Sci USA 78:3760–3764
Ito H, Fukuda Y, Murata K, Kimura A (1983) Transformation of intact yeast cells treated with alkali cations. J Bacteriol 153:163–168
Jayaram M, Li Y-Y, Broach JR (1983) The yeast plasmid 2 μ circle encodes components required for its high copy propogation. Cell 34:95–104
Jayaram M, Sutton A, Broach JR (1985) Properties of REP3: a cis-acting locus required for stable propagation of the Saccharomyces cerevisiae plasmid 2 μm circle. Mol Cell Biol 5:2466–2475
Kikuchi Y (1983) Yeast plasmid requires a cis-acting locus and two plasmid proteins for its stable maintenance. Cell 35:487–493
Livingston DM, Kupfer DM (1977) Control of Saccharomyces cerevisiae 2 micron DNA replication by cell division cycle genes that control nuclear DNA replication. J Mol Biol 116:249–260
Maniatis T, Fritsch EF, Sambrook J (1982) Molecular cloning. A laboratory manual. Cold Spring harbor Laboratory, Cold Spring Harbor, New York
Messing J (1983) New M13 vectors for cloning. Methods in Enzymology 101:20–78
Murray AW, Szostak JW (1983) Pedigree analysis of plasmid segregation in yeast. Cell 34:961–970
Ogura T, Hiraga S (1982) Partition mechanism of F plasmid: Two plasmid gene-encoded products and a cis-acting region are involved in partitioning. Cell 32:351–360
Orr-Weaver TL, Szostak JW, Rothstein RS (1981) Yeast transformation: A model system for the study of recombination. Proc Natl Acad Sci USA 78:6354–6358
Petes TD, Williamson DH (1975) Replicating circular DNA molecules in yeast. Cell 4:249–253
Sinclair JH, Stevens BJ, Sanghavi P, Rabinowitz M (1967) Mitochondrial-satellite and circular DNA filaments in yeast. Science 156:1234–1237
Southern EM (1975) Detection of specific sequences amongst DNA fragments separated by gel electrophoresis. J Mol Biol 98:503–517
Stinchcomb DT, Struhl K, Davis RW (1979) Isolation and characterisation of a yeast chromosomal replicator. Nature 282:39–43
Struhl K, Stinchcomb DT, Sherer S, Davis RW (1979) High frequency transformation of yeast: Autonomous replication of hybrid DNA molecules. Proc Natl Acad Sci USA 76:1035–1039
Veit BE, Fangman WL (1985) Chromatin organisation of the Saccharomyces cerevisiae 2 μm plasmid depends on plasmid-encoded products. Mol Cell Biol 5:2190–2196
Wahl GM, Stern M, Stark GR (1979) Efficient transfer of large DNA fragments from agarose gels to diazobenzyloxymethylpaper and rapid hybridisation by using dextran sulphate. Proc Natl Acad Sci USA 76:3683–3687
Zakian VA, Brewer BJ, Fangman WL (1979) Replication of each copy of the yeast 2 micron DNA plasmid occurs during the S phase. Cell 17:923–934
Author information
Authors and Affiliations
Additional information
Communicated by C.P. Hollenberg
Rights and permissions
About this article
Cite this article
Cashmore, A.M., Albury, M.S., Hadfield, C. et al. Genetic analysis of partitioning functions encoded by the 2 μm circle of Saccharomyces cerevisiae . Mol Gen Genet 203, 154–162 (1986). https://doi.org/10.1007/BF00330397
Received:
Issue Date:
DOI: https://doi.org/10.1007/BF00330397