PMCCPMCCPMCC

Search tips
Search criteria 

Advanced

 
Logo of jbacterPermissionsJournals.ASM.orgJournalJB ArticleJournal InfoAuthorsReviewers
 
J Bacteriol. 1997 December; 179(23): 7497–7506.
PMCID: PMC179702

The 2microm-plasmid-encoded Rep1 and Rep2 proteins interact with each other and colocalize to the Saccharomyces cerevisiae nucleus.

Abstract

The efficient partitioning of the 2microm plasmid of Saccharomyces cerevisiae at cell division requires two plasmid-encoded proteins (Rep1p and Rep2p) and a cis-acting locus, REP3 (STB). By using protein hybrids containing fusions of the Rep proteins to green fluorescent protein (GFP), we show here that fluorescence from GFP-Rep1p or GFP-Rep2p is almost exclusively localized in the nucleus in a cir+ strain. Nuclear localization of GFP-Rep1p and GFP-Rep2p, though discernible, is less efficient in a cir(0) host. GFP-Rep2p or GFP-Rep1p is able to promote the stability of a 2microm circle-derived plasmid harboring REP1 or REP2, respectively, in a cir(0) background. Under these conditions, fluorescence from GFP-Rep2p or GFP-Rep1p is concentrated within the nucleus, as is the case in cir+ cells. This characteristic nuclear accumulation is not dependent on the expression of the FLP or RAF1 gene of the 2microm circle. Nuclear colocalization of Rep1p and Rep2p is consistent with the hypothesis that the two proteins directly or indirectly interact to form a functional bipartite or high-order protein complex. Immunoprecipitation experiments as well as baiting assays using GST-Rep hybrid proteins suggest a direct interaction between Rep1p and Rep2p which, in principle, may be modulated by other yeast proteins. Furthermore, these assays provide evidence for Rep1p-Rep1p and Rep2p-Rep2p associations as well. The sum of these interactions may be important in controlling the effective cellular concentration of the Rep1p-Rep2p complex.

Full Text

The Full Text of this article is available as a PDF (7.3M).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.
  • Chalfie M, Tu Y, Euskirchen G, Ward WW, Prasher DC. Green fluorescent protein as a marker for gene expression. Science. 1994 Feb 11;263(5148):802–805. [PubMed]
  • Dobson MJ, Yull FE, Molina M, Kingsman SM, Kingsman AJ. Reconstruction of the yeast 2 micron plasmid partitioning mechanism. Nucleic Acids Res. 1988 Jul 25;16(14B):7103–7117. [PMC free article] [PubMed]
  • Doyle T, Botstein D. Movement of yeast cortical actin cytoskeleton visualized in vivo. Proc Natl Acad Sci U S A. 1996 Apr 30;93(9):3886–3891. [PubMed]
  • Futcher AB. Copy number amplification of the 2 micron circle plasmid of Saccharomyces cerevisiae. J Theor Biol. 1986 Mar 21;119(2):197–204. [PubMed]
  • Futcher AB. The 2 micron circle plasmid of Saccharomyces cerevisiae. Yeast. 1988 Mar;4(1):27–40. [PubMed]
  • Futcher AB, Cox BS. Maintenance of the 2 microns circle plasmid in populations of Saccharomyces cerevisiae. J Bacteriol. 1983 May;154(2):612–622. [PMC free article] [PubMed]
  • Hadfield C, Mount RC, Cashmore AM. Protein binding interactions at the STB locus of the yeast 2 microns plasmid. Nucleic Acids Res. 1995 Mar 25;23(6):995–1002. [PMC free article] [PubMed]
  • Hartley JL, Donelson JE. Nucleotide sequence of the yeast plasmid. Nature. 1980 Aug 28;286(5776):860–865. [PubMed]
  • Jayaram M, Sutton A, Broach JR. Properties of REP3: a cis-acting locus required for stable propagation of the Saccharomyces cerevisiae plasmid 2 microns circle. Mol Cell Biol. 1985 Sep;5(9):2466–2475. [PMC free article] [PubMed]
  • Jayaram M, Sumida S, Young LJ. Inducible expression of REP1 causes inducible expression of the 2 micron circle stability system. Curr Genet. 1986;11(2):85–91. [PubMed]
  • Jayaram M, Li YY, Broach JR. The yeast plasmid 2mu circle encodes components required for its high copy propagation. Cell. 1983 Aug;34(1):95–104. [PubMed]
  • Kikuchi Y. Yeast plasmid requires a cis-acting locus and two plasmid proteins for its stable maintenance. Cell. 1983 Dec;35(2 Pt 1):487–493. [PubMed]
  • Laemmli UK. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. [PubMed]
  • Ma X, Ehrhardt DW, Margolin W. Colocalization of cell division proteins FtsZ and FtsA to cytoskeletal structures in living Escherichia coli cells by using green fluorescent protein. Proc Natl Acad Sci U S A. 1996 Nov 12;93(23):12998–13003. [PubMed]
  • Murray AW, Szostak JW. Pedigree analysis of plasmid segregation in yeast. Cell. 1983 Oct;34(3):961–970. [PubMed]
  • Murray JA, Cesareni G. Functional analysis of the yeast plasmid partition locus STB. EMBO J. 1986 Dec 1;5(12):3391–3399. [PubMed]
  • Murray JA, Cesareni G, Argos P. Unexpected divergence and molecular coevolution in yeast plasmids. J Mol Biol. 1988 Apr 5;200(3):601–607. [PubMed]
  • Murray JA, Scarpa M, Rossi N, Cesareni G. Antagonistic controls regulate copy number of the yeast 2 mu plasmid. EMBO J. 1987 Dec 20;6(13):4205–4212. [PubMed]
  • Petes TD, Williamson DH. A novel structural form of the 2 micron plasmid of the yeast Saccharomyces cerevisiae. Yeast. 1994 Oct;10(10):1341–1345. [PubMed]
  • Reynolds AE, Murray AW, Szostak JW. Roles of the 2 microns gene products in stable maintenance of the 2 microns plasmid of Saccharomyces cerevisiae. Mol Cell Biol. 1987 Oct;7(10):3566–3573. [PMC free article] [PubMed]
  • Russo FD, Scherson I, Broach JR. Direct simulation of yeast 2-microns circle plasmid amplification. J Theor Biol. 1992 Apr 7;155(3):369–385. [PubMed]
  • Sigurdson DC, Gaarder ME, Livingston DM. Characterization of the transmission during cytoductant formation of the 2 micrometers DNA plasmid from Saccharomyces. Mol Gen Genet. 1981;183(1):59–65. [PubMed]
  • Som T, Armstrong KA, Volkert FC, Broach JR. Autoregulation of 2 micron circle gene expression provides a model for maintenance of stable plasmid copy levels. Cell. 1988 Jan 15;52(1):27–37. [PubMed]
  • Utatsu I, Sakamoto S, Imura T, Toh-e A. Yeast plasmids resembling 2 micron DNA: regional similarities and diversities at the molecular level. J Bacteriol. 1987 Dec;169(12):5537–5545. [PMC free article] [PubMed]
  • Veit BE, Fangman WL. Copy number and partition of the Saccharomyces cerevisiae 2 micron plasmid controlled by transcription regulators. Mol Cell Biol. 1988 Nov;8(11):4949–4957. [PMC free article] [PubMed]
  • Volkert FC, Broach JR. Site-specific recombination promotes plasmid amplification in yeast. Cell. 1986 Aug 15;46(4):541–550. [PubMed]
  • Volkert FC, Wilson DW, Broach JR. Deoxyribonucleic acid plasmids in yeasts. Microbiol Rev. 1989 Sep;53(3):299–317. [PMC free article] [PubMed]
  • Waddle JA, Karpova TS, Waterston RH, Cooper JA. Movement of cortical actin patches in yeast. J Cell Biol. 1996 Mar;132(5):861–870. [PMC free article] [PubMed]
  • Wu LC, Fisher PA, Broach JR. A yeast plasmid partitioning protein is a karyoskeletal component. J Biol Chem. 1987 Jan 15;262(2):883–891. [PubMed]
  • Zakian VA, Brewer BJ, Fangman WL. Replication of each copy of the yeast 2 micron DNA plasmid occurs during the S phase. Cell. 1979 Aug;17(4):923–934. [PubMed]

Articles from Journal of Bacteriology are provided here courtesy of American Society for Microbiology (ASM)