Logo of molcellbPermissionsJournals.ASM.orgJournalMCB ArticleJournal InfoAuthorsReviewers
Mol Cell Biol. Jun 1997; 17(6): 3261–3271.
PMCID: PMC232179

A lesion in the DNA replication initiation factor Mcm10 induces pausing of elongation forks through chromosomal replication origins in Saccharomyces cerevisiae.

Abstract

We describe a new minichromosome maintenance factor, Mcm10, and show that this essential protein is involved in the initiation of DNA replication in Saccharomyces cerevisiae. The mcm10 mutant has an autonomously replicating sequence-specific minichromosome maintenance defect and arrests at the nonpermissive temperature with dumbbell morphology and 2C DNA content. Mcm10 is a nuclear protein that physically interacts with several members of the MCM2-7 family of DNA replication initiation factors. Cloning and sequencing of the MCM10 gene show that it is identical to DNA43, a gene identified independently for its putative role in replicating DNA. Two-dimensional DNA gel analysis reveals that the mcm10-1 lesion causes a dramatic reduction in DNA replication initiation at chromosomal origins, including ORI1 and ORI121. Interestingly, the mcm10-1 lesion also causes replication forks to pause during elongation through these same loci. This novel phenotype suggests a unique role for the Mcm10 protein in the initiation of DNA synthesis at replication origins.

Full Text

The Full Text of this article is available as a PDF (2.0M).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.
  • Althoefer H, Schleiffer A, Wassmann K, Nordheim A, Ammerer G. Mcm1 is required to coordinate G2-specific transcription in Saccharomyces cerevisiae. Mol Cell Biol. 1995 Nov;15(11):5917–5928. [PMC free article] [PubMed]
  • Bell SP, Stillman B. ATP-dependent recognition of eukaryotic origins of DNA replication by a multiprotein complex. Nature. 1992 May 14;357(6374):128–134. [PubMed]
  • Boeke JD, LaCroute F, Fink GR. A positive selection for mutants lacking orotidine-5'-phosphate decarboxylase activity in yeast: 5-fluoro-orotic acid resistance. Mol Gen Genet. 1984;197(2):345–346. [PubMed]
  • Brewer BJ, Fangman WL. The localization of replication origins on ARS plasmids in S. cerevisiae. Cell. 1987 Nov 6;51(3):463–471. [PubMed]
  • Brewer BJ, Fangman WL. A replication fork barrier at the 3' end of yeast ribosomal RNA genes. Cell. 1988 Nov 18;55(4):637–643. [PubMed]
  • Chen Y, Hennessy KM, Botstein D, Tye BK. CDC46/MCM5, a yeast protein whose subcellular localization is cell cycle-regulated, is involved in DNA replication at autonomously replicating sequences. Proc Natl Acad Sci U S A. 1992 Nov 1;89(21):10459–10463. [PMC free article] [PubMed]
  • Chong JP, Mahbubani HM, Khoo CY, Blow JJ. Purification of an MCM-containing complex as a component of the DNA replication licensing system. Nature. 1995 Jun 1;375(6530):418–421. [PubMed]
  • Chong JP, Thömmes P, Blow JJ. The role of MCM/P1 proteins in the licensing of DNA replication. Trends Biochem Sci. 1996 Mar;21(3):102–106. [PubMed]
  • Dershowitz A, Newlon CS. The effect on chromosome stability of deleting replication origins. Mol Cell Biol. 1993 Jan;13(1):391–398. [PMC free article] [PubMed]
  • Deshpande AM, Newlon CS. The ARS consensus sequence is required for chromosomal origin function in Saccharomyces cerevisiae. Mol Cell Biol. 1992 Oct;12(10):4305–4313. [PMC free article] [PubMed]
  • Deshpande AM, Newlon CS. DNA replication fork pause sites dependent on transcription. Science. 1996 May 17;272(5264):1030–1033. [PubMed]
  • Diffley JF, Cocker JH, Dowell SJ, Rowley A. Two steps in the assembly of complexes at yeast replication origins in vivo. Cell. 1994 Jul 29;78(2):303–316. [PubMed]
  • Dijkwel PA, Vaughn JP, Hamlin JL. Mapping of replication initiation sites in mammalian genomes by two-dimensional gel analysis: stabilization and enrichment of replication intermediates by isolation on the nuclear matrix. Mol Cell Biol. 1991 Aug;11(8):3850–3859. [PMC free article] [PubMed]
  • Elble R. A simple and efficient procedure for transformation of yeasts. Biotechniques. 1992 Jul;13(1):18–20. [PubMed]
  • Feinberg AP, Vogelstein B. A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity. Anal Biochem. 1983 Jul 1;132(1):6–13. [PubMed]
  • Ferguson BM, Brewer BJ, Reynolds AE, Fangman WL. A yeast origin of replication is activated late in S phase. Cell. 1991 May 3;65(3):507–515. [PubMed]
  • Fields S, Song O. A novel genetic system to detect protein-protein interactions. Nature. 1989 Jul 20;340(6230):245–246. [PubMed]
  • Fox CA, Loo S, Dillin A, Rine J. The origin recognition complex has essential functions in transcriptional silencing and chromosomal replication. Genes Dev. 1995 Apr 15;9(8):911–924. [PubMed]
  • Gavin KA, Hidaka M, Stillman B. Conserved initiator proteins in eukaryotes. Science. 1995 Dec 8;270(5242):1667–1671. [PubMed]
  • Gibson SI, Surosky RT, Tye BK. The phenotype of the minichromosome maintenance mutant mcm3 is characteristic of mutants defective in DNA replication. Mol Cell Biol. 1990 Nov;10(11):5707–5720. [PMC free article] [PubMed]
  • Gossen M, Pak DT, Hansen SK, Acharya JK, Botchan MR. A Drosophila homolog of the yeast origin recognition complex. Science. 1995 Dec 8;270(5242):1674–1677. [PubMed]
  • Greenfeder SA, Newlon CS. Replication forks pause at yeast centromeres. Mol Cell Biol. 1992 Sep;12(9):4056–4066. [PMC free article] [PubMed]
  • Hennessy KM, Lee A, Chen E, Botstein D. A group of interacting yeast DNA replication genes. Genes Dev. 1991 Jun;5(6):958–969. [PubMed]
  • Hutter KJ, Eipel HE. Flow cytometric determinations of cellular substances in algae, bacteria, moulds and yeasts. Antonie Van Leeuwenhoek. 1978;44(3-4):269–282. [PubMed]
  • Kearsey SE, Maiorano D, Holmes EC, Todorov IT. The role of MCM proteins in the cell cycle control of genome duplication. Bioessays. 1996 Mar;18(3):183–190. [PubMed]
  • Kilmartin JV, Adams AE. Structural rearrangements of tubulin and actin during the cell cycle of the yeast Saccharomyces. J Cell Biol. 1984 Mar;98(3):922–933. [PMC free article] [PubMed]
  • Kobayashi T, Hidaka M, Nishizawa M, Horiuchi T. Identification of a site required for DNA replication fork blocking activity in the rRNA gene cluster in Saccharomyces cerevisiae. Mol Gen Genet. 1992 Jun;233(3):355–362. [PubMed]
  • Koonin EV. A common set of conserved motifs in a vast variety of putative nucleic acid-dependent ATPases including MCM proteins involved in the initiation of eukaryotic DNA replication. Nucleic Acids Res. 1993 Jun 11;21(11):2541–2547. [PMC free article] [PubMed]
  • Kubota Y, Mimura S, Nishimoto S, Takisawa H, Nojima H. Identification of the yeast MCM3-related protein as a component of Xenopus DNA replication licensing factor. Cell. 1995 May 19;81(4):601–609. [PubMed]
  • Lei M, Kawasaki Y, Tye BK. Physical interactions among Mcm proteins and effects of Mcm dosage on DNA replication in Saccharomyces cerevisiae. Mol Cell Biol. 1996 Sep;16(9):5081–5090. [PMC free article] [PubMed]
  • Liang C, Weinreich M, Stillman B. ORC and Cdc6p interact and determine the frequency of initiation of DNA replication in the genome. Cell. 1995 Jun 2;81(5):667–676. [PubMed]
  • Linskens MH, Huberman JA. Organization of replication of ribosomal DNA in Saccharomyces cerevisiae. Mol Cell Biol. 1988 Nov;8(11):4927–4935. [PMC free article] [PubMed]
  • Madine MA, Khoo CY, Mills AD, Laskey RA. MCM3 complex required for cell cycle regulation of DNA replication in vertebrate cells. Nature. 1995 Jun 1;375(6530):421–424. [PubMed]
  • Maine GT, Sinha P, Tye BK. Mutants of S. cerevisiae defective in the maintenance of minichromosomes. Genetics. 1984 Mar;106(3):365–385. [PMC free article] [PubMed]
  • Maiorano D, Van Assendelft GB, Kearsey SE. Fission yeast cdc21, a member of the MCM protein family, is required for onset of S phase and is located in the nucleus throughout the cell cycle. EMBO J. 1996 Feb 15;15(4):861–872. [PMC free article] [PubMed]
  • Marahrens Y, Stillman B. A yeast chromosomal origin of DNA replication defined by multiple functional elements. Science. 1992 Feb 14;255(5046):817–823. [PubMed]
  • Moir D, Botstein D. Determination of the order of gene function in the yeast nuclear division pathway using cs and ts mutants. Genetics. 1982 Apr;100(4):565–577. [PMC free article] [PubMed]
  • Moir D, Stewart SE, Osmond BC, Botstein D. Cold-sensitive cell-division-cycle mutants of yeast: isolation, properties, and pseudoreversion studies. Genetics. 1982 Apr;100(4):547–563. [PMC free article] [PubMed]
  • Passmore S, Elble R, Tye BK. A protein involved in minichromosome maintenance in yeast binds a transcriptional enhancer conserved in eukaryotes. Genes Dev. 1989 Jul;3(7):921–935. [PubMed]
  • Passmore S, Maine GT, Elble R, Christ C, Tye BK. Saccharomyces cerevisiae protein involved in plasmid maintenance is necessary for mating of MAT alpha cells. J Mol Biol. 1988 Dec 5;204(3):593–606. [PubMed]
  • Rhode PR, Elsasser S, Campbell JL. Role of multifunctional autonomously replicating sequence binding factor 1 in the initiation of DNA replication and transcriptional control in Saccharomyces cerevisiae. Mol Cell Biol. 1992 Mar;12(3):1064–1077. [PMC free article] [PubMed]
  • Rose MD, Novick P, Thomas JH, Botstein D, Fink GR. A Saccharomyces cerevisiae genomic plasmid bank based on a centromere-containing shuttle vector. Gene. 1987;60(2-3):237–243. [PubMed]
  • Solomon NA, Wright MB, Chang S, Buckley AM, Dumas LB, Gaber RF. Genetic and molecular analysis of DNA43 and DNA52: two new cell-cycle genes in Saccharomyces cerevisiae. Yeast. 1992 Apr;8(4):273–289. [PubMed]
  • Takahashi K, Yamada H, Yanagida M. Fission yeast minichromosome loss mutants mis cause lethal aneuploidy and replication abnormality. Mol Biol Cell. 1994 Oct;5(10):1145–1158. [PMC free article] [PubMed]
  • Treisman JE, Follette PJ, O'Farrell PH, Rubin GM. Cell proliferation and DNA replication defects in a Drosophila MCM2 mutant. Genes Dev. 1995 Jul 15;9(14):1709–1715. [PubMed]
  • Tye BK. The MCM2-3-5 proteins: are they replication licensing factors? Trends Cell Biol. 1994 May;4(5):160–166. [PubMed]
  • Walker SS, Malik AK, Eisenberg S. Analysis of the interactions of functional domains of a nuclear origin of replication from Saccharomyces cerevisiae. Nucleic Acids Res. 1991 Nov 25;19(22):6255–6262. [PMC free article] [PubMed]
  • Yan H, Gibson S, Tye BK. Mcm2 and Mcm3, two proteins important for ARS activity, are related in structure and function. Genes Dev. 1991 Jun;5(6):944–957. [PubMed]
  • Yan H, Merchant AM, Tye BK. Cell cycle-regulated nuclear localization of MCM2 and MCM3, which are required for the initiation of DNA synthesis at chromosomal replication origins in yeast. Genes Dev. 1993 Nov;7(11):2149–2160. [PubMed]

Articles from Molecular and Cellular Biology are provided here courtesy of American Society for Microbiology (ASM)

Formats:

Related citations in PubMed

See reviews...See all...

Cited by other articles in PMC

See all...

Links

  • Gene
    Gene
    Gene links
  • GEO Profiles
    GEO Profiles
    Related GEO records
  • HomoloGene
    HomoloGene
    HomoloGene links
  • MedGen
    MedGen
    Related information in MedGen
  • Pathways + GO
    Pathways + GO
    Pathways, annotations and biological systems (BioSystems) that cite the current article.
  • Protein
    Protein
    Published protein sequences
  • PubMed
    PubMed
    PubMed citations for these articles
  • Taxonomy
    Taxonomy
    Related taxonomy entry
  • Taxonomy Tree
    Taxonomy Tree

Recent Activity

Your browsing activity is empty.

Activity recording is turned off.

Turn recording back on

See more...