Skip to main content
SpringerLink
Log in

The human autoantigen MCP1 is required during early stages of DNA replication

  • Published:
Chromosome Research Aims and scope Submit manuscript

Abstract

Metaphase chromosome protein 1 (MCP1) is a nuclear autoantigen that is associated with condensed chromosomes throughout mitosis. During interphase, this antigen shows a speckle distribution in the nucleus, excluding the nucleolus. Additionally, MCP1 binds tightly to the scaffold/matrix component of nuclei and isolated chromosomes. In order to determine the in-vivo localization of the antigen, we have expressed MCP1 fused to EGFP in tissue culture cells. The results demonstrate that MCP1 is located in the nucleus during interphase and during mitosis associates tightly to condensed chromosomes. Furthermore, microinjection of specific antibody confirms these results. We have used a specific monoclonal antibody (mAb 402) against MCP1 to assess the function of this antigen during cell cycle progression. HeLa and Ptk-2 cells that were microinjected into the nucleus and/or cytoplasm at G1/S and very early S phase were not able to progress and complete DNA replication. However, injection of mAb 402 at mid or late S phase does not prevent completion of DNA replication and subsequent progression into mitosis. Microinjection of mAb 402 in Ptk-2 cells synchronized in mitosis did not interfere with progression of mitosis and cells divided. Our results suggest that MCP1 is required at the G1/S transition and during early S phase.

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

  • Balczon R (1993) Autoantibodies as probes in cell and molecular biology. Proc Soc Exp Biol Med 204: 138–154.

    PubMed  CAS  Google Scholar 

  • Bernat RL, Borisy GG, Rothfield NF, Earnshaw WC (1990) Injection of anticentromere antibodies in interphase cells disrupts events required for chromosome movement at mitosis. J Cell Biol 111: 1519–1533.

    Article  PubMed  CAS  Google Scholar 

  • Blow JJ, Tada S (2000) A new check on issuing the licence. Nature 404: 560–561.

    Article  PubMed  CAS  Google Scholar 

  • Bronze-da-Rocha E, Machado C, Staines NA, Sunkel CE (1992) Systemic lupus erythematosus murine monoclonal DNA-binding antibodies recognize cytoplasmic and nuclear phosphorylated antigens that display cell cycle redistribution in HEp-2 cells. Immunology 77: 582–591.

    PubMed  CAS  Google Scholar 

  • Bronze-da-Rocha E, Catita JA, Sunkel CE (1998) Molecular cloning of metaphase chromosome protein 1 (MCP1), a novel human autoantigen that associates with condensed chromosomes during mitosis. Chromosome Res 6: 85–95.

    Article  PubMed  CAS  Google Scholar 

  • Cohen PL, Eisenberg R (1991) Lpr and gld: single gene models of systemic autoimmunity and lymphoproliferative disease. Annu Rev Immunol 9: 234–269.

    Article  Google Scholar 

  • Coleman TR, Carpenter PB, Dunphy WG (1996) The Xenopus Cdc6 protein is essential for the initiation of a single round of DNA replication in cell-free extracts. Cell 87: 53–63.

    Article  PubMed  CAS  Google Scholar 

  • Coué M, Kearsey SE, Méchali M (1996) Chromatin binding, nuclear-localization and phosphorylation of Xenopus cdc21 are cell-cycle dependent and associated with the control of initiation of DNA replication. EMBO J 15 861–872.

    Google Scholar 

  • Fairman MP (1990) DNA polimerase δ/PCNA: actions and interactions. J Cell Sci 95: 1–4.

    PubMed  CAS  Google Scholar 

  • Fakan S, Hancock R (1974) Localization of newly-synthesized DNA in a mammalian cell as visualized by high resolution autoradiography. Exp Cell Res 83: 95–102.

    Article  PubMed  CAS  Google Scholar 

  • Ferreira J, Paolella G, Ramos C, Lamond, AI (1997) Spatial organization of large-scale chromatin domains in the nucleus: a magnified view of single chromosome territories. J Cell Biol 139: 1597–1610.

    Article  PubMed  CAS  Google Scholar 

  • Fox MH, Arndt-Jovin DJ, Jovin TM, Baumann PH, Robert-Nicoud M (1991) Spatial and temporal distribution of DNA replication sites localized by immunofluorescence and confocal microscopy in mouse fibroblasts. J Cell Sci 99: 247–253.

    PubMed  Google Scholar 

  • Görlich D, Kutay U (1999) Transport between the cell nucleus and the cytoplasm. Annu Rev Cell Dev Biol 15: 607–660.

    Article  PubMed  Google Scholar 

  • Gottlieb E, Steitz JA (1989a) The RNA binding protein La influences both the accuracy and the efficiency of RNA polimerase III transcription in vitro. EMBO J 8: 841–850.

    PubMed  CAS  Google Scholar 

  • Gottlieb E, Steitz JA (1989b) Function of the mammalian La protein: evidence for its action in transcription termination by RNA polimerase III. EMBO J 8: 851–861.

    PubMed  CAS  Google Scholar 

  • Harlow E, Lane D (1988) Antibodies-A Laboratory Manual, 1st Edn. New York: Cold Spring Harbor Laboratory Press.

    Google Scholar 

  • Hunt T, Sherr CJ (1994) Cell multiplication. Curr Opin Cell Biol 6: 833–835.

    Article  CAS  Google Scholar 

  • Isenberg DA, Ehrenstein MR, Longhurst C, Kalsi JK (1994) The origin, sequence, structure, and consequences of developing anti-DNA antibodies. Arthritis Rheum 37: 169–180.

    PubMed  CAS  Google Scholar 

  • Jackson DA, Pombo A (1998) Replication clusters are stable units of chromosome structure: evidence that nuclear organization contributes to the efficient activation and propagation of S phase in human cells. J Cell Biol 140: 1285–1295.

    Article  PubMed  CAS  Google Scholar 

  • Kaffman A, O'Shea EK (1999) Regulation of nuclear localization: a key to a door. Annu Rev Cell Dev Biol 15: 291–339.

    Article  PubMed  CAS  Google Scholar 

  • Kelman Z (1997) PCNA: structure, functions and interactions. Oncogene 14: 629–640.

    Article  PubMed  CAS  Google Scholar 

  • King RW, Deshaies RJ, Petters J-M, Kirschner MW (1996) How proteolysis drives cell cycle. Science 274: 1652–1659.

    Article  PubMed  CAS  Google Scholar 

  • Koike M, Awaji T, Kataoka M et al. (1999) Differential sub-cellular localization of DNA-dependent protein kinase components Ku and DNA-PKcs during mitosis. J Cell Sci 112: 4031–4039.

    PubMed  CAS  Google Scholar 

  • Laemmli UK (1970) Cleavage of structural protein during assembly of the head of bacteriophage T4. Nature 227: 680–685.

    Article  PubMed  CAS  Google Scholar 

  • Lake R, Morgan A, Staines N (1987) Production of monoclonal antibodies. Soc Appl Microbiol Tech Ser 24: 1–19.

    Google Scholar 

  • Ma H, Samarabandu J, Devdhar RS et al. (1998) Spatial and temporal dynamics of DNA replication sites in mammalian cells. J Cell Biol 143: 1415–1425.

    Article  PubMed  CAS  Google Scholar 

  • Mahbubani HM, Chong JPJ, Chevalier S, Thömmes P, Blow JJ (1997) Cell cycle regulation of the replication licensing system: involvement of a Cdk-dependent inhibitor. J Cell Biol 136: 125–135.

    Article  PubMed  CAS  Google Scholar 

  • Maiorano D, Moreau J, Méchali M (2000) XCDT1 is required for the assembly of prereplicative complexes in Xenopus laevis. Nature 404: 622–625.

    Article  PubMed  CAS  Google Scholar 

  • Morgan A, Buchanan RRC, Lew AM, Oslen I, Staines NA (1985a) Five groups of antigenic determinants on DNA identified by monoclonal antobodies from (NZB x NZW)F1 and MRL/Mp-lpr/lpr mice. Immunology 55: 75–83.

    PubMed  CAS  Google Scholar 

  • Morgan A, Isenberg DA, Naparstek et al. (1985b) Shared idiotypes are expressed on mouse and human anti-DNA autoantibodies. Immunology 56: 393–399.

    PubMed  CAS  Google Scholar 

  • Nasmyth K (1996) Viewpoint: putting the cell cycle in order. Science 274: 1643–1645.

    Article  PubMed  CAS  Google Scholar 

  • O'Keefe RT, Henderson SC, Spector DL (1992) Dynamic organization of DNA replication in mammalian cell nuclei: spatially and temporally defined replication of chromosome-specific α-satellite sequences. J Cell Biol 116: 1095–1110.

    Article  PubMed  Google Scholar 

  • Rowles A, Blow JJ (1997) Chromatin proteins involved in the initiation of DNA replication. Curr Opin Gen Dev 7: 152–157.

    Article  CAS  Google Scholar 

  • Steinberg AD, Gourley MF, Klinman DM, Tsokos GC, Scott DE, Krieg AM (1991) Systemic lupus erythematosus. Ann Intern Med 115: 548–559.

    PubMed  CAS  Google Scholar 

  • Tan EM (1989) Antinuclear antibodies: diagnostic markers for autoimmune diseases and probes for cell biology. Adv Immunol 44: 93–151.

    Article  PubMed  CAS  Google Scholar 

  • Tan EM (1991) Autoantibodies in pathology and cell biology. Cell 67: 841–842.

    Article  PubMed  CAS  Google Scholar 

  • Tan EM (1994) Autoimmunity and apoptosis. J Exp Med 179: 1083–1086.

    Article  PubMed  CAS  Google Scholar 

  • Towbin H, Staehelin T, Gordon J (1979) Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci USA 76: 4350–4354.

    Article  PubMed  CAS  Google Scholar 

  • Vega-Salas EE, Salas PJI (1996) Cell cycle related behaviour of a chromosomal scaffold protein in MDCK epithelial cells. Chromosoma 104: 321–331.

    PubMed  CAS  Google Scholar 

  • Watts RA, Ravirajan CT, Staines NA, Isenberg DA (1990) A human fetal monoclonal DNA-binding antibody shares idiotypes with fetal and adult murine monoclonal DNA-binding antibodies. Immunology 69: 348–354.

    PubMed  CAS  Google Scholar 

  • Zachariae W (1999) Progression into and out of mitosis. Curr Opin Cell Biol 11: 708–716.

    Article  PubMed  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Laboratório de Genética Molecular da Mitose, Instituto de Biologia Molecular e Celular, da Universidade do Porto, Rua do Campo Alegre, 823, 4150-180, Porto, Portugal.

    Elsa Bronze-da-Rocha, Ana Nóvoa & Claudio E. Sunkel (Largo Professor)

  2. Laboratório de Bioquímica da Faculdade de Farmácia, da Universidade do Porto, Rua Anibal Cunha, 164, 4050-047, Porto, Portugal

    Elsa Bronze-da-Rocha

  3. Instituto de Histologia e Embriologia da Faculdade de Medicina, da Universidade de Lisboa, 1699, Lisboa Codex, Portugal

    Celso Cunha & Maria do Carmo-Fonseca

  4. Infection & Immunity Research Group, Division of Life Sciences, King's College London, University of London, London, SE1 8WA, UK

    Norman A. Staines

  5. Instituto de Ciências Biomédicas Abel Salazar, da Universidade do Porto, Abel Salazar, 2, 4099-003, Porto, Portugal

    Claudio E. Sunkel (Largo Professor)

Authors
  1. Elsa Bronze-da-Rocha
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Ana Nóvoa
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Celso Cunha
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Maria do Carmo-Fonseca
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Norman A. Staines
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Claudio E. Sunkel
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Bronze-da-Rocha, E., Nóvoa, A., Cunha, C. et al. The human autoantigen MCP1 is required during early stages of DNA replication. Chromosome Res 8, 699–711 (2000). https://doi.org/10.1023/A:1012097704355

Download citation

  • Issue Date:

  • DOI: https://doi.org/10.1023/A:1012097704355

Search

Navigation