Skip to main content
SpringerLink
Log in

cDNA cloning and characterisation of a maize homologue of the MCM proteins required for the initiation of DNA replication

  • Original Paper
  • Published:
Molecular and General Genetics MGG Aims and scope Submit manuscript

Abstract

A central question in cell cycle regulation is how DNA replication is initiated and executed only once in each cell cycle. The cell cycle-regulated assembly of specific initiation protein complexes at chromosomal origins appears to specify the initial sites and timing of DNA replication, and to restrict this process to only one round in the somatic cell cycle. Among the enzymes involved in origin activation, the MCM proteins play a conserved key role. In particular, MCM3 homologues have been shown to be components of the DNA replication licensing activity in yeast and vertebrates. In spite of our detailed knowledge of the regulation of the initiation of DNA synthesis in yeast, there is virtually no information available on the molecules involved in origin activation in higher plants. We have isolated a cDNA from maize root apices, termedROA (Replication Origin Activator), encoding a protein which shares a high degree of homology with the MCM3 subfamily of MCM proteins. Analysis of gene organisation by Southern blotting shows 2–4 copies per haploid genome of closely relatedROA sequences and the presence of further less related sequences in a multigene family. The steady-state levels ofROA mRNA are under developmental control, being relatively high in proliferative tissues such as the root apex, the developing cob and the coleoptile, and are strongly correlated with that of the histone H4 transcript.In situ hybridisation analysis in the root apex reveals thatROA mRNA expression is limited to specific subpopulations of cycling cells, which is typical of cell cycle-regulated expression. The isolation of nearly identical sequences from barley andArabidopsis by the polymerase chain reaction indicates that MCM-related proteins are conserved in higher plants.

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

Access this article

Log in via an institution

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

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  • Aves SJ, Bryant JA (1993) The molecular genetics and biochemistry of DNA replication. In: Ormrod JC, Francis D (eds) Molecular and cell biology of the plant cell cycle. Kluwer Academic Publishers, Amsterdam, pp 45–56

    Google Scholar 

  • Bell SP, Stillman B (1992) ATP-dependent recognition of eukaryotic origins of DNA replication by a multiprotein complex. Nature 357:128–134

    Google Scholar 

  • Blow JJ, Laskey RA (1988) A role for the nuclear envelope in controlling DNA replication within the cell cycle. Nature 332:546–548

    Google Scholar 

  • Bucci S, Ragghianti M, Nardi I, Bellini M, Mancino G, Lacroix, JC (1993) Identification of an amphibian oocyte nuclear protein as a candidate for a role in embryonic DNA replication. Int J Dev Biol 37:509–517

    Google Scholar 

  • Bussereau F, Mallet L, Gaillon L, Jacquet M (1993) A 12.8 kb segment, on the right arm of chromosome II fromSaccharomyces cerevisiae including part of theDUR1,2 gene, contains five putative new genes. Yeast 9:797–806

    Google Scholar 

  • Chan CSM, Tye B-K (1980) Autonomously replicating sequences inSaccharomyces cerevisiae. Proc Natl Acad Sci USA 77:6329–6333

    Google Scholar 

  • Chaubet N, Clément B, Phillips G, Gigot C (1991) Organ-specific expression of different histone H3 and H4 gene subfamilies in developing and adult maize. Plant Mol Biol 17:935–940

    Google Scholar 

  • Chen Y, Hennessy KM, Botstein D, Tye B-K (1992) CDC46/MCM5, a yeast protein whose subcellular localization is cell-cycle regulated, is involved in DNA replication at ARSs. Proc Natl Acad Sci USA 89:10459–10463

    Google Scholar 

  • Chong JPJ, Mahbubani HM, Khoo C-Y, Blow JJ (1995) Purification of an MCM-containing complex as a component of the DNA replication licensing system. Nature 375:418–421

    Google Scholar 

  • Coverly D, Laskey RA (1994) Regulation of eukaryotic DNA replication. Annu Rev Biochem 63:745–776

    Google Scholar 

  • Coxon A, Maundrell K, Kearsey SE (1992) Fission yeastcdc21 + belongs to a family of proteins involved in an early step of chromosome replication. Nucleic Acids Res 20:5571–5577

    Google Scholar 

  • Dalton S, Whitbread L (1995) Cell cycle-regulated nuclear import and export of Cdc47, a protein essential for initiation of DNA replication in budding yeast. Proc Natl Acad Sci USA 92:2514–2518

    Google Scholar 

  • D'Amato F (1984) Role of polyploidy in reproductive organs and tissues. In: Johri BM (ed.) Embryology of angiosperms. Springer-Verlag, New York, pp 519–566

    Google Scholar 

  • Devereux J, Haeberli P, Smithies O (1984) A comprehensive set of sequence analysis programs for the VAX. Nucleic Acids Res 12:387–395

    Google Scholar 

  • Elble R (1992) A simple and efficient procedure for transformation of yeasts. BioTechniques 13:18–20

    Google Scholar 

  • Fobert PR, Coen EC, Murphy GJP, Doonan JH (1994) Patterns of cell division revealed by transcriptional regulation of genes during the cell cycle in plants. EMBO J 13:616–624

    Google Scholar 

  • Forsburg S, Nurse P (1994) The fission yeastcdc19 + gene encodes a member of the MCM family of replication proteins. J Cell Sci 107:2779–2788

    Google Scholar 

  • Frohman MA, Dush MK, Martin GR (1988) Rapid production of full length cDNAs from rare transcripts: amplification using a single gene specific oligonucleotide primer. Proc Natl Acad Sci USA 85:8998–9002

    Google Scholar 

  • Gibson SI, Surosky RT, Tye B-K (1990) The phenotype of the minichromosome mutantmcm3 is characteristic of mutants defective in DNA replication. Mol Cell Biol 10:5707–5720

    Google Scholar 

  • Guthrie C, Fink GR (eds) (1991) Guide to yeast genetics and molecular biology. Methods Enzymol 194

  • Hennessy KM, Clark CD, Botstein D (1990) Subcellular localization of yeast Cdc46 varies with the cell cycle. Genes Dev 4:2252–2263

    Google Scholar 

  • Hennessy KM, Lee A, Chen E, Botstein D (1991) A group of interacting yeast DNA replication genes. Genes Dev 5:958–969

    Google Scholar 

  • Hu B, Burkhart R, Schulte D, Musahl C, Knippers R (1993) The P1 family: a new class of nuclear mammalian proteins related to the yeast Mcm replication proteins. Nucleic Acids Res 21:5289–5293

    Google Scholar 

  • Huberman JA (1995) Prokaryotic and eukaryotic replicons. Cell 82:535–542

    Google Scholar 

  • Johnston LH, Lowndes NF (1992) Cell cycle control of DNA synthesis in budding yeast. Nucleic Acids Res 20:2403–2410

    Google Scholar 

  • Kimura H, Nozaki N, Sugimoto K (1994) DNA polymerase α-associated protein P1, a murine homolog of yeast MCM3, changes its intranuclear distribution during the DNA synthetic period. EMBO J 13:4311–4320

    Google Scholar 

  • Koonin EV (1993) 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 21:2541–2547

    Google Scholar 

  • Kubota Y, Satoru M, Nishimoto S, Takisawa H, Nojima H (1995) Identification of the yeast MCM3-related protein as a component ofXenopus DNA replication licensing factor. Cell 81:601–609

    Google Scholar 

  • Li JJ, Herskowitz I (1993) Isolation ofORC6, a component of the yeast origin recognition complex by a one-hybrid system. Science 262:1870–1874

    Google Scholar 

  • Logemann J, Schell J, Willmitzer L (1987) Improved method for the isolation of RNA from plant tissues. Anal Biochem 163:16–20

    Google Scholar 

  • Madine MA, Khoo C-Y, Millis AD, Laskey RA (1995) MCM3 complex required for cell cycle regulation of DNA replication in vertebrate cells. Nature 375:421–424

    Google Scholar 

  • Miyake S, Okishio N, Samejima I, Hiraoka Y, Toda T, Saitoh I, Yanagida M (1993) Fission yeast genesnda1 + andnda4 +, mutations of which lead to S-phase block, chromatin alteration and Ca2+ suppression, are members of theCDC46/MCM2 family. Mol Biol Cell 4:1003–1015

    Google Scholar 

  • Moreno S, Nurse P (1990) Substrates for p34cdc2:In vivo veritas? Cell 61:549–551

    Google Scholar 

  • Nagl W (1978) Endopolyploidy and polyteny in differentiation and evolution. Elsevier/North-Holland, Amsterdam

    Google Scholar 

  • Nurse P (1990) Universal control mechanism regulating onset of M-phase. Nature 344:503–508

    Google Scholar 

  • Osley MA (1991) The regulation of histone synthesis in the cell cycle. Annu Rev Biochem 60:827–886

    Google Scholar 

  • Phillips G, Chauvet M, Chaubote ME, Gigot C (1986) Genomic organisation and nucleotide sequences of two corn histone genes. Gene 42:225–229

    Google Scholar 

  • Rao PN, Johnson RN (1970) Mammalian cell fusion: studies on the regulation of DNA synthesis and mitosis. Nature 225:159–164

    Google Scholar 

  • Rost B, Sander C (1993) Improved prediction of protein secondary structure by use of sequence profiles and neural networks. Proc Natl Acad Sci USA 90:7558–7562

    Google Scholar 

  • Ruiz-Avila L, Burgess SR, Stiefel V, Ludevid MD, Puigdomenech P (1992) Accumulation of cell wall hydroxyproline-rich glycoprotein mRNA is an early event in maize embryo cell differentiation. Proc Natl Acad Sci USA 89:2414–2418

    Google Scholar 

  • Sabelli PA, Shewry PR (1991) Characterisation and organisation of gene families at theGli-1 loci of bread and durum wheat by restriction fragment analysis. Theor Appl Genet 83:209–216

    Google Scholar 

  • Sabelli PA, Shewry PR (1993) Nucleic acid blotting and hybridisation. In: Bryant JA (ed) Methods in plant biochemistry, vol 10. Academic Press, London, pp 79–100

    Google Scholar 

  • Saraste M, Sibbald PR, Wittinghofer A (1990) The P-loop—a common motif in ATP- and GTP-binding proteins. Trends Biochem Sci 15:430–434

    Google Scholar 

  • Schulte D, Burkhart R, Musahl C, Hu B, Schlatterer C, Hameister H, Knippers R (1995) Expression, phosphorylation and nuclear localisation of the human P1 protein, a homologue of the yeast Mcm3 replication protein. J Cell Sci 108:1381–1389

    Google Scholar 

  • Schweizer L, Yerk-Davis GL, Phillips RL, Srienc F, Jones RJ (1995) Dynamics of maize endosperm development and DNA endoreduplication. Proc Natl Acad Sci USA 92:7070–7074

    Google Scholar 

  • Sikorski RS, Hieter P (1989) A system of shuttle vectors and yeast host strains designed for efficient manipulation of DNA inSaccharomyces cerevisiae. Genetics 122:19–27

    Google Scholar 

  • Springer PS, McCombie WR, Sundarasen V, Martienssen RA (1995) Gene trap tagging ofPROLIFERA, an essentialMCM2-3-5-like gene inArabidopsis. Science 268:877–880

    Google Scholar 

  • Starborg, M, Brundell E, Gell K, Larsson C, White I, Daneholt B, Höög C (1995) A murine replication protein accumulates temporarily in the heterochromatic regions of nuclei prior to initiation of DNA replication. J Cell Sci 108:927–934

    Google Scholar 

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

    Google Scholar 

  • Takahashi K, Yamada H, Yanagida M (1994) Fission yeast minichromosome loss mutantsmis cause lethal aneuploidy and replication abnormality. Mol Biol Cell 5:1145–1158

    Google Scholar 

  • Thömmes P, Fett R, Schray B, Burkhart R, Barnes M, Kennedy C, Brown NC, Knippers R (1992) Properties of the nuclear P1 protein, a mammalian homologue of the yeast Mcm3 replication protein. Nucleic Acids Res 20:1069–1074

    Google Scholar 

  • Todorov IT, Peppertok R, Philipova RN, Kearsey SE, Ansorge W, Werner D (1994) A human nuclear protein with sequence homology to a family of early S phase proteins is required for entry into S phase and for cell division. J Cell Sci 107:253–265

    Google Scholar 

  • Toyn JH, Toone WM, Morgan BA, Johnston LH (1995) The activation of DNA replication in yeast. Trends Biochem Sci 20:70–73

    Google Scholar 

  • Tye B-K (1994) The MCM2-3-5 proteins are they replication licensing factors? Trends Cell Biol 4:160–166

    Google Scholar 

  • Walker JE, Saraste M, Runswick MJ, Gay NJ (1982) Distantly related sequences in the α- and β-subunits of ATP synthase, myosin, kinases and other ATP-requiring enzymes and a common nucleotide binding fold. EMBO J 1:945–951

    Google Scholar 

  • Wilson R, Ainscough R, Anderson K, Baynes C, Berks M, Bonfield J, Burton J, Connell M, Copsey T, Cooper J, Coulson A, Craxton M, Dear S, Du Z, Burbin R, Favello A, Fraser A, Fulton L, Gardner A, Green P, Hawkins T, Hillier L, Jier M, Johnston L, Jones M, Kershaw J, Kirsten J, Laisster N, Latreille P, Lightning J, Lloyd C, Mortimore B, O'Callaghan M, Parsons J, Percy C, Rifken L, Roopra A, Saunders D, Shownkeen R, Sims M, Smaldon N, Smith A, Smith M, Sonnhammer E, Staden R, Sulston J, Thierry-Mieg J, Thomas K, Vaudin M, Vaughan K, Waterston R, Watson A, Weinstock L, Wilkinson-Sproat J, Wohldman P (1994) 2.2 Mb of contiguous nucleotide sequence from chromosome III ofC. elegans. Nature 368:32–38

    Google Scholar 

  • Yan H, Gibson S, Tye B-K (1991) Mcm2 and Mcm3, two proteins important for ARS activity, are related in structure and function. Genes Dev 5:944–957

    Google Scholar 

  • Yan H, Merchant AM, Tye B-K (1993) Cell cycle-regulated nuclear localisation of MCM2 and MCM3, which are required for the initiation of DNA synthesis at chromosomal replication origins in yeast. Genes Dev 7:2149–2160

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. IACR-Long Ashton Research Station, Department of Agricultural Sciences, University of Bristol, BS18 9AF, Bristol, UK

    Paolo A. Sabelli, Shirley R. Burgess & Peter R. Shewry

  2. Institute of Molecular and Cell Biology, National University of Singapore, 10 Kent Ridge Crescent, 0511, Singapore, Republic of Singapore

    Paolo A. Sabelli & Anil K. Kush

  3. Section of Biochemistry, Molecular and Cell Biology, Cornell University, 14853, Ithaca, New York, USA

    Michael R. Young

Authors
  1. Paolo A. Sabelli
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Shirley R. Burgess
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Anil K. Kush
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Michael R. Young
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Peter R. Shewry
    View author publications

    You can also search for this author in PubMed Google Scholar

Additional information

Communicated by A. Kondorosi

Rights and permissions

Reprints and permissions

About this article

Cite this article

Sabelli, P.A., Burgess, S.R., Kush, A.K. et al. cDNA cloning and characterisation of a maize homologue of the MCM proteins required for the initiation of DNA replication. Molec. Gen. Genet. 252, 125–136 (1996). https://doi.org/10.1007/BF02173212

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF02173212

Key words

Search

Navigation