DNA topoisomerase II alpha is the major chromosome protein recognized by the mitotic phosphoprotein antibody MPM-2 - PubMed (original) (raw)
DNA topoisomerase II alpha is the major chromosome protein recognized by the mitotic phosphoprotein antibody MPM-2
S Taagepera et al. Proc Natl Acad Sci U S A. 1993.
Abstract
We have determined that the major mitotic phosphoprotein in chromosomes recognized by the antiphosphoprotein antibody MPM-2 is the 170-kDa isoform of topoisomerase II (topo II), the isoform predominant in proliferating cells. As a prerequisite to making this discovery, it was necessary to develop protocols to protect chromosomal proteins from dephosphorylation during cell extraction and chromosome isolation procedures. Immunofluorescence analysis of the large chromosomes prepared from Indian Muntjac cells revealed colocalization of MPM-2 and anti-topo II antibodies to the chromosomal centromeres and to the axial regions of the chromosomal arms. For biochemical fractionation studies, large quantities of chromosomes from the P388D1 mouse lymphocyte cell line were isolated and treated to remove DNA and histone proteins. Immunoblot and immunoprecipitation experiments with this chromosome scaffold fraction identified the major MPM-2-reactive phosphoprotein to be DNA topo II. Using a panel of anti-peptide antibodies specific to the isoforms of topo II, we determined that the major phosphoprotein recognized by MPM-2 is the 170-kDa isoform of topo II, topo II alpha. The 180-kDa isoform, topo II beta, present in the isolated chromosomes in much smaller quantities, is also recognized by MPM-2. The mitotic phosphorylation of the topo II proteins may be critical for proper chromosome condensation and segregation.
Similar articles
- DNA topoisomerase II distribution in mouse preimplantation embryos.
St Pierre J, Wright DJ, Rowe TC, Wright SJ. St Pierre J, et al. Mol Reprod Dev. 2002 Mar;61(3):335-46. doi: 10.1002/mrd.10016. Mol Reprod Dev. 2002. PMID: 11835579 - Cell-cycle-regulated localization of tyrosine and threonine phosphoepitopes at the kinetochores of mitotic chromosomes.
Taagepera S, Campbell MS, Gorbsky GJ. Taagepera S, et al. Exp Cell Res. 1995 Nov;221(1):249-60. doi: 10.1006/excr.1995.1373. Exp Cell Res. 1995. PMID: 7589252 - The distribution of topoisomerase II on mammalian chromosomes.
Sumner AT. Sumner AT. Chromosome Res. 1996 Jan;4(1):5-14. doi: 10.1007/BF02254938. Chromosome Res. 1996. PMID: 8653270 - Topoisomerase II: untangling its contribution at the centromere.
Porter AC, Farr CJ. Porter AC, et al. Chromosome Res. 2004;12(6):569-83. doi: 10.1023/B:CHRO.0000036608.91085.d1. Chromosome Res. 2004. PMID: 15289664 Review. - Untangling the role of DNA topoisomerase II in mitotic chromosome structure and function.
Warburton PE, Earnshaw WC. Warburton PE, et al. Bioessays. 1997 Feb;19(2):97-9. doi: 10.1002/bies.950190203. Bioessays. 1997. PMID: 9046238 Review.
Cited by
- Molecular mechanisms of anthracycline cardiovascular toxicity.
Narezkina A, Narayan HK, Zemljic-Harpf AE. Narezkina A, et al. Clin Sci (Lond). 2021 May 28;135(10):1311-1332. doi: 10.1042/CS20200301. Clin Sci (Lond). 2021. PMID: 34047339 Free PMC article. Review. - Mitotic chromosomes.
Paulson JR, Hudson DF, Cisneros-Soberanis F, Earnshaw WC. Paulson JR, et al. Semin Cell Dev Biol. 2021 Sep;117:7-29. doi: 10.1016/j.semcdb.2021.03.014. Epub 2021 Apr 6. Semin Cell Dev Biol. 2021. PMID: 33836947 Free PMC article. Review. - Histone H2A phosphorylation recruits topoisomerase IIα to centromeres to safeguard genomic stability.
Zhang M, Liang C, Chen Q, Yan H, Xu J, Zhao H, Yuan X, Liu J, Lin S, Lu W, Wang F. Zhang M, et al. EMBO J. 2020 Feb 3;39(3):e101863. doi: 10.15252/embj.2019101863. Epub 2019 Nov 26. EMBO J. 2020. PMID: 31769059 Free PMC article. - A Monoclonal Antibody to M-Phase Phosphoprotein 1/Kinesin-Like Protein KIF20B.
Fritzler MJ, Brown RD, Zhang M. Fritzler MJ, et al. Monoclon Antib Immunodiagn Immunother. 2019 Aug;38(4):162-170. doi: 10.1089/mab.2019.0016. Epub 2019 Jul 16. Monoclon Antib Immunodiagn Immunother. 2019. PMID: 31260385 Free PMC article. - The Impact of the C-Terminal Region on the Interaction of Topoisomerase II Alpha with Mitotic Chromatin.
Antoniou-Kourounioti M, Mimmack ML, Porter ACG, Farr CJ. Antoniou-Kourounioti M, et al. Int J Mol Sci. 2019 Mar 12;20(5):1238. doi: 10.3390/ijms20051238. Int J Mol Sci. 2019. PMID: 30871006 Free PMC article.
References
- EMBO J. 1986 May;5(5):1003-10 - PubMed
- Cancer Res. 1991 Aug 1;51(15):3951-7 - PubMed
- Proc Natl Acad Sci U S A. 1977 Nov;74(11):4937-41 - PubMed
- Eur J Biochem. 1980 Oct;111(1):189-97 - PubMed
- Cell. 1982 May;29(1):171-81 - PubMed
Publication types
MeSH terms
Substances
LinkOut - more resources
Full Text Sources
Research Materials