Skip to main content
Log in

Characterization of a Mis12 homologue in Arabidopsis thaliana

  • Published:
Chromosome Research Aims and scope Submit manuscript

Abstract

The centromere/kinetochore represents an important complex on chromosomes that contains a large number of proteins and facilitates accurate chromosome segregation during cell division. Fission yeast Mis12 and its human homologue hMis12 have been identified as essential kinetochore components. Although homologues have been suggested to exist in plants, their function remains to be determined. In this study the full-length cDNA of the Mis12 homologue from Arabidopsis thaliana (AtMIS12) was successfully cloned by RACE-and RT-PCR and the DNA sequence determined. The 238 amino acid sequence deduced from the cDNA contains two conserved blocks and a coiled-coil motif, despite the poor overall similarity to fission yeast and human Mis12. The antibody raised against a partial peptide of AtMIS12 recognized a 27-kDa protein corresponding to the predicted molecular weight. Immunofluorescence labeling using the antibody revealed that AtMIS12 localizes at centromeric regions, like the centromeric histone H3 variant HTR12, throughout the cell cycle. These results indicate that AtMIS12 is a constitutive component of Arabidopsis kinetochores.

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

Access this article

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

Instant access to the full article PDF.

Similar content being viewed by others

Reference

  • Ahmad K, Henikoff S (2002) Histone H3 variants specify modes of chromatin assembly. Proc Natl Acad Sci USA 99 (Suppl 4): 16477–16484.

    PubMed  CAS  Google Scholar 

  • Amor DJ, Bentley K, Ryan J et al. (2004) Human centromere repositioning “in progress”. Proc Natl Acad Sci USA 101: 6542–6547.

    Article  PubMed  CAS  Google Scholar 

  • Dawe RK, Reed LM, Yu HG et al. (1999) A maize homolog of mammalian CENPC is a constitutive component of the inner kinetochore. Plant Cell 11: 1227–1238.

    Article  PubMed  CAS  Google Scholar 

  • Fukagawa T, Pendon C, Morris J, Brown W (1999) CENP-C is necessary but not sufficient to induce formation of a functional centromere. EMBO J 18: 4196–4209.

    Article  PubMed  CAS  Google Scholar 

  • Fukagawa T, Mikami Y, Nishihashi A et al. (2001) CENP-H, a constitutive centromere component, is required for centromere targeting of CENP-C in vertebrate cells. EMBO J 20: 4603–4617.

    Article  PubMed  CAS  Google Scholar 

  • Goshima G, Saitoh S, Yanagida M (1999) Proper metaphase spindle length is determined by centromere proteins Mis12 and Mis6 required for faithful chromosome segregation. Genes Dev 13: 1664–1677.

    PubMed  CAS  Google Scholar 

  • Goshima G, Kiyomitsu T, Yoda K, Yanagida M (2003) Human centromere chromatin protein hMis12, essential for equal segregation, is independent of CENP-A loading pathway. J Cell Biol 160: 25–39.

    Article  PubMed  CAS  Google Scholar 

  • Henikoff S, Ahmad K, Malik HS (2001) The centromere paradox: stable inheritance with rapidly evolving DNA. Science 293: 1098–1102.

    Article  PubMed  CAS  Google Scholar 

  • Howman EV, Fowler KJ, Newson AJ et al. (2000) Early disruption of centromeric chromatin organization in centromere protein A (Cenpa) null mice. Proc Natl Acad Sci USA 97: 1148–1153.

    Article  PubMed  CAS  Google Scholar 

  • Liu ST, Hittle JC, Jablonski SA et al. (2003) Human CENP-I specifies localization of CENP-F, MAD1 and MAD2 to kinetochores and is essential for mitosis. Nat Cell Biol 5: 341–345.

    Article  PubMed  CAS  Google Scholar 

  • Mathur J, Szabados L, Schaefer S et al. (1998) Gene identification with sequenced T-DNA tags generated by transformation of Arabidopsis cell suspension. Plant J 13: 707–716.

    Article  PubMed  CAS  Google Scholar 

  • Murata M (2002) Telomeres and centromeres in plants. Curr Genom 3: 527–538.

    CAS  Google Scholar 

  • Nagaki K, Talbert PB, Zhong CX et al. (2003) Chromatin immunoprecipitation reveals that the 180-bp satellite repeat Is the key functional DNA element of Arabidopsis thaliana centromeres. Genetics 163: 1221–1225.

    PubMed  CAS  Google Scholar 

  • Nagaki K, Cheng Z, Ouyang, S. et al. (2004) Sequencing of a rice centromere uncovers active genes. Nat Genet 36: 138–145.

    Article  PubMed  CAS  Google Scholar 

  • Nishihashi A, Haraguchi T, Hiraoka Y et al. (2002) CENP-I is essential for centromere function in vertebrate cells. Dev Cell 2: 463–476.

    Article  PubMed  CAS  Google Scholar 

  • Ogura Y, Shibata F, Sato H, Murata M (2004) Characterization of a CENP-C homolog in Arabidopsis thaliana. Genes Genet Syst 79: 139–144.

    Article  PubMed  CAS  Google Scholar 

  • Shibata F, Murata M (2004) Differential localization of the centromere-specific proteins in the major centromeric satellite of Arabidopsis thaliana. J Cell Sci 117: 2963–2970.

    Article  PubMed  CAS  Google Scholar 

  • Sugata N, Li S, Earnshaw WC et al. (2000) Human CENP-H multimers colocalize with CENP-A and CENP-C at active centromere–kinetochore complexes. Hum Mol Genet 9: 2919–2926.

    Article  PubMed  CAS  Google Scholar 

  • Takahashi K, Chen ES, Yanagida M (2000) Requirement of Mis6 centromere connector for localizing a CENP-A-like protein in fission yeast. Science 288: 2215–2219.

    Article  PubMed  CAS  Google Scholar 

  • Talbert PB, Masuelli R, Tyagi AP et al. (2002) Centromeric localization and adaptive evolution of an Arabidopsis histone H3 variant. Plant Cell 14: 1053–1066.

    Article  PubMed  CAS  Google Scholar 

  • Talbert PB, Bryson TD, Henikoff S (2004) Adaptive evolution of centromere proteins in plants and animals. J Biol 3: 18.

    Article  PubMed  Google Scholar 

  • Zhong CX, Marshall JB, Topp C et al. (2002) Centromeric retroelements and satellites interact with maize kinetochore protein CENH3. Plant Cell 14: 2825–2836.

    PubMed  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Minoru Murata.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Sato, H., Shibata, F. & Murata, M. Characterization of a Mis12 homologue in Arabidopsis thaliana. Chromosome Res 13, 827–834 (2005). https://doi.org/10.1007/s10577-005-1016-3

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10577-005-1016-3

Key words

Navigation