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J Cell Biol. 1985 May 1; 100(5): 1706–1715.
PMCID: PMC2113886

Topoisomerase II is a structural component of mitotic chromosome scaffolds


We have obtained a polyclonal antibody that recognizes a major polypeptide component of chicken mitotic chromosome scaffolds. This polypeptide migrates in SDS PAGE with Mr 170,000. Indirect immunofluorescence and subcellular fractionation experiments confirm that it is present in both mitotic chromosomes and interphase nuclei. Two lines of evidence suggest that this protein is DNA topoisomerase II, an abundant nuclear enzyme that controls DNA topological states: anti-scaffold antibody inhibits the strand-passing activity of DNA topoisomerase II; and both anti-scaffold antibody and an independent antibody raised against purified bovine topoisomerase II recognize identical partial proteolysis fragments of the 170,000-mol-wt scaffold protein in immunoblots. Our results suggest that topoisomerase II may be an enzyme that is also a structural protein of interphase nuclei and mitotic chromosomes.

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Selected References

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  • Rattner JB, Goldsmith M, Hamkalo BA. Chromatin organization during meiotic prophase of Bombyx mori. Chromosoma. 1980;79(2):215–224. [PubMed]
  • Rattner JB, Goldsmith MR, Hamkalo BA. Chromosome organization during male meiosis in Bombyx mori. Chromosoma. 1981;82(3):341–351. [PubMed]
  • Bahr GF. Human chromosome fibers. Considerations of DNA-protein packing and of looping patterns. Exp Cell Res. 1970 Sep;62(1):39–49. [PubMed]
  • Adolph KW, Cheng SM, Laemmli UK. Role of nonhistone proteins in metaphase chromosome structure. Cell. 1977 Nov;12(3):805–816. [PubMed]
  • Paulson JR, Laemmli UK. The structure of histone-depleted metaphase chromosomes. Cell. 1977 Nov;12(3):817–828. [PubMed]
  • Laemmli UK, Cheng SM, Adolph KW, Paulson JR, Brown JA, Baumbach WR. Metaphase chromosome structure: the role of nonhistone proteins. Cold Spring Harb Symp Quant Biol. 1978;42(Pt 1):351–360. [PubMed]
  • Adolphs KW, Cheng SM, Paulson JR, Laemmli UK. Isolation of a protein scaffold from mitotic HeLa cell chromosomes. Proc Natl Acad Sci U S A. 1977 Nov;74(11):4937–4941. [PMC free article] [PubMed]
  • Rattner JB, Branch A, Hamkalo BA. Electron microscopy of whole mount metaphase chromosomes. Chromosoma. 1975 Nov 11;52(4):329–338. [PubMed]
  • Labhart P, Koller T, Wunderli H. Involvement of higher order chromatin structures in metaphase chromosome organization. Cell. 1982 Aug;30(1):115–121. [PubMed]
  • Okada TA, Comings DE. A search for protein cores in chromosomes: is the scaffold an artifact? Am J Hum Genet. 1980 Nov;32(6):814–832. [PMC free article] [PubMed]
  • Earnshaw WC, Laemmli UK. Architecture of metaphase chromosomes and chromosome scaffolds. J Cell Biol. 1983 Jan;96(1):84–93. [PMC free article] [PubMed]
  • Earnshaw WC, Heck MM. Localization of topoisomerase II in mitotic chromosomes. J Cell Biol. 1985 May;100(5):1716–1725. [PMC free article] [PubMed]
  • Benyajati C, Worcel A. Isolation, characterization, and structure of the folded interphase genome of Drosophila melanogaster. Cell. 1976 Nov;9(3):393–407. [PubMed]
  • Cook PR, Brazell IA. Spectrofluorometric measurement of the binding of ethidium to superhelical DNA from cell nuclei. Eur J Biochem. 1978 Mar 15;84(2):465–477. [PubMed]
  • Igó-Kemenes T, Zachau HG. Domains in chromatin structure. Cold Spring Harb Symp Quant Biol. 1978;42(Pt 1):109–118. [PubMed]
  • Adolph KW. Organization of chromosomes in HeLa cells: isolation of histone-depleted nuclei and nuclear scaffolds. J Cell Sci. 1980 Apr;42:291–304. [PubMed]
  • Vogelstein B, Pardoll DM, Coffey DS. Supercoiled loops and eucaryotic DNA replicaton. Cell. 1980 Nov;22(1 Pt 1):79–85. [PubMed]
  • Lebkowski JS, Laemmli UK. Evidence for two levels of DNA folding in histone-depleted HeLa interphase nuclei. J Mol Biol. 1982 Apr 5;156(2):309–324. [PubMed]
  • Hyde JE. Expansion of chicken erythrocyte nuclei upon limited micrococcal nuclease digestion. Correlation with higher order chromatin structure. Exp Cell Res. 1982 Jul;140(1):63–70. [PubMed]
  • Sundin O, Varshavsky A. Terminal stages of SV40 DNA replication proceed via multiply intertwined catenated dimers. Cell. 1980 Aug;21(1):103–114. [PubMed]
  • Sundin O, Varshavsky A. Arrest of segregation leads to accumulation of highly intertwined catenated dimers: dissection of the final stages of SV40 DNA replication. Cell. 1981 Sep;25(3):659–669. [PubMed]
  • Liu LF. DNA topoisomerases--enzymes that catalyse the breaking and rejoining of DNA. CRC Crit Rev Biochem. 1983;15(1):1–24. [PubMed]
  • DiNardo S, Voelkel K, Sternglanz R. DNA topoisomerase II mutant of Saccharomyces cerevisiae: topoisomerase II is required for segregation of daughter molecules at the termination of DNA replication. Proc Natl Acad Sci U S A. 1984 May;81(9):2616–2620. [PMC free article] [PubMed]
  • Uemura T, Yanagida M. Isolation of type I and II DNA topoisomerase mutants from fission yeast: single and double mutants show different phenotypes in cell growth and chromatin organization. EMBO J. 1984 Aug;3(8):1737–1744. [PMC free article] [PubMed]
  • Earnshaw WC, Rothfield N. Identification of a family of human centromere proteins using autoimmune sera from patients with scleroderma. Chromosoma. 1985;91(3-4):313–321. [PubMed]
  • Halligan BD, Edwards KA, Liu LF. Purification and characterization of a type II DNA topoisomerase from bovine calf thymus. J Biol Chem. 1985 Feb 25;260(4):2475–2482. [PubMed]
  • Earnshaw WC, Halligan N, Cooke C, Rothfield N. The kinetochore is part of the metaphase chromosome scaffold. J Cell Biol. 1984 Jan;98(1):352–357. [PMC free article] [PubMed]
  • Towbin H, Staehelin T, Gordon J. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci U S A. 1979 Sep;76(9):4350–4354. [PMC free article] [PubMed]
  • Laskey RA, Mills AD. Quantitative film detection of 3H and 14C in polyacrylamide gels by fluorography. Eur J Biochem. 1975 Aug 15;56(2):335–341. [PubMed]
  • Lebkowski JS, Laemmli UK. Non-histone proteins and long-range organization of HeLa interphase DNA. J Mol Biol. 1982 Apr 5;156(2):325–344. [PubMed]
  • Miller KG, Liu LF, Englund PT. A homogeneous type II DNA topoisomerase from HeLa cell nuclei. J Biol Chem. 1981 Sep 10;256(17):9334–9339. [PubMed]
  • Liu LF, Miller KG. Eukaryotic DNA topoisomerases: two forms of type I DNA topoisomerases from HeLa cell nuclei. Proc Natl Acad Sci U S A. 1981 Jun;78(6):3487–3491. [PMC free article] [PubMed]
  • Liu LF, Davis JL, Calendar R. Novel topologically knotted DNA from bacteriophage P4 capsids: studies with DNA topoisomerases. Nucleic Acids Res. 1981 Aug 25;9(16):3979–3989. [PMC free article] [PubMed]
  • Berrios M, Filson AJ, Blobel G, Fisher PA. A 174-kilodalton ATPase/dATPase polypeptide and a glycoprotein of apparently identical molecular weight are common but distinct components of higher eukaryotic nuclear structural protein subfractions. J Biol Chem. 1983 Nov 10;258(21):13384–13390. [PubMed]
  • Takagi N, Sasaki M. A phylogenetic study of bird karyotypes. Chromosoma. 1974 May 21;46(1):91–120. [PubMed]
  • Cozzarelli NR. DNA gyrase and the supercoiling of DNA. Science. 1980 Feb 29;207(4434):953–960. [PubMed]
  • Gellert M. DNA topoisomerases. Annu Rev Biochem. 1981;50:879–910. [PubMed]
  • Duguet M, Lavenot C, Harper F, Mirambeau G, De Recondo AM. DNA topoisomerases from rat liver: physiological variations. Nucleic Acids Res. 1983 Feb 25;11(4):1059–1075. [PMC free article] [PubMed]
  • Hadlaczky G, Sumner AT, Ross A. Protein-depleted chromosomes. II. Experiments concerning the reality of chromosome scaffolds. Chromosoma. 1981;81(4):557–567. [PubMed]
  • Marsden MP, Laemmli UK. Metaphase chromosome structure: evidence for a radial loop model. Cell. 1979 Aug;17(4):849–858. [PubMed]
  • Buongiorno-Nardelli M, Micheli G, Carri MT, Marilley M. A relationship between replicon size and supercoiled loop domains in the eukaryotic genome. Nature. 1982 Jul 1;298(5869):100–102. [PubMed]
  • Stalder J, Larsen A, Engel JD, Dolan M, Groudine M, Weintraub H. Tissue-specific DNA cleavages in the globin chromatin domain introduced by DNAase I. Cell. 1980 Jun;20(2):451–460. [PubMed]
  • Lawson GM, Knoll BJ, March CJ, Woo SL, Tsai MJ, O'Malley BW. Definition of 5' and 3' structural boundaries of the chromatin domain containing the ovalbumin multigene family. J Biol Chem. 1982 Feb 10;257(3):1501–1507. [PubMed]
  • IZAWA M, ALLEFREY VG, MIRSKY AE. The relationship between RNA synthesis and loop structure in lampbrush chromosomes. Proc Natl Acad Sci U S A. 1963 Apr;49:544–551. [PMC free article] [PubMed]
  • Callan HG. The Croonian Lecture, 1981. Lampbrush chromosomes. Proc R Soc Lond B Biol Sci. 1982 Mar 22;214(1197):417–448. [PubMed]
  • Luchnik AN, Bakayev VV, Zbarsky IB, Georgiev GP. Elastic torsional strain in DNA within a fraction of SV40 minichromosomes: relation to transcriptionally active chromatin. EMBO J. 1982;1(11):1353–1358. [PMC free article] [PubMed]
  • Harland RM, Weintraub H, McKnight SL. Transcription of DNA injected into Xenopus oocytes is influenced by template topology. Nature. 1983 Mar 3;302(5903):38–43. [PubMed]
  • Nordheim A, Pardue ML, Lafer EM, Möller A, Stollar BD, Rich A. Antibodies to left-handed Z-DNA bind to interband regions of Drosophila polytene chromosomes. Nature. 1981 Dec 3;294(5840):417–422. [PubMed]
  • Nordheim A, Lafer EM, Peck LJ, Wang JC, Stollar BD, Rich A. Negatively supercoiled plasmids contain left-handed Z-DNA segments as detected by specific antibody binding. Cell. 1982 Dec;31(2 Pt 1):309–318. [PubMed]
  • Weintraub H. A dominant role for DNA secondary structure in forming hypersensitive structures in chromatin. Cell. 1983 Apr;32(4):1191–1203. [PubMed]
  • Ryoji M, Worcel A. Chromatin assembly in Xenopus oocytes: in vivo studies. Cell. 1984 May;37(1):21–32. [PubMed]
  • Steck TR, Drlica K. Bacterial chromosome segregation: evidence for DNA gyrase involvement in decatenation. Cell. 1984 Apr;36(4):1081–1088. [PubMed]
  • Williamson DH, Fennell DJ. The use of fluorescent DNA-binding agent for detecting and separating yeast mitochondrial DNA. Methods Cell Biol. 1975;12:335–351. [PubMed]

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