Mitotic chromatin condensation in vitro using somatic cell extracts and nuclei with variable levels of endogenous topoisomerase II

doi:10.1083/jcb.111.6.2839

This Article

Full Text (PDF, 3297K)

Alert me when this article is cited

Citation Map

Services

Email this article

Similar articles in this journal

Similar articles in PubMed

Alert me to new content in the JCB

Download to citation manager

Citing Articles

Citing Articles via HighWire

Citing Articles via CrossRef

Citing Articles via Google Scholar

Google Scholar

Articles by Wood, E. R.

Articles by Earnshaw, W. C.

Search for Related Content

PubMed

PubMed Citation

Articles by Wood, E. R.

Articles by Earnshaw, W. C.

Pubmed/NCBI databases

Substance via MeSH

Social Bookmarking

What's this?

ARTICLES

Mitotic chromatin condensation in vitro using somatic cell extracts and nuclei with variable levels of endogenous topoisomerase II

ER Wood and WC Earnshaw
Department of Cell Biology and Anatomy, Johns Hopkins University School of Medicine, Baltimore, Maryland 21205.

We report the development of a new method for producing mitotic extracts from tissue culture cells. These extracts reproducibly promote the condensation of chromatin in vitro when incubated with purified interphase nuclei. This condensation reaction is not species specific, since nuclei from chicken, human, and hamster cell lines all undergo chromatin condensation upon incubation with the extract. We have used this extract to investigate the role of DNA topoisomerase II (topo II) in the chromosome condensation process. Chromatin condensation does not require the presence of soluble topo II in the mitotic extract. However, the extent of formation of discrete chromosome-like structures correlates with the level of endogenous topo II present in the interphase nuclei. Our results further suggest that chromatin condensation in this extract may involve two processes: chromatin compaction and resolution into discrete chromosomes.
Add to CiteULike CiteULike Add to Complore Complore Add to Connotea Connotea Add to Del.icio.us Del.icio.us Add to Digg Digg Facebook Add to Reddit Reddit Add to Technorati Technorati Twitter What's this?

This article has been cited by other articles:

Roca, J. (2009). Topoisomerase II: a fitted mechanism for the chromatin landscape. Nucleic Acids Res 37: 721-730 [Abstract] [Full Text]
Ageichik, A. V., Samejima, K., Kaufmann, S. H., Earnshaw, W. C. (2007). Genetic Analysis of the Short Splice Variant of the Inhibitor of Caspase-activated DNase (ICAD-S) in Chicken DT40 Cells. J. Biol. Chem. 282: 27374-27382 [Abstract] [Full Text]
Martins, R. P., Krawetz, S. A. (2007). Decondensing the protamine domain for transcription. Proc. Natl. Acad. Sci. USA 104: 8340-8345 [Abstract] [Full Text]
Zhang, A., Lyu, Y. L., Lin, C.-P., Zhou, N., Azarova, A. M., Wood, L. M., Liu, L. F. (2006). A Protease Pathway for the Repair of Topoisomerase II-DNA Covalent Complexes. J. Biol. Chem. 281: 35997-36003 [Abstract] [Full Text]
Lyu, Y. L., Lin, C.-P., Azarova, A. M., Cai, L., Wang, J. C., Liu, L. F. (2006). Role of Topoisomerase II{beta} in the Expression of Developmentally Regulated Genes. Mol. Cell. Biol. 26: 7929-7941 [Abstract] [Full Text]
Andrews, C. A., Vas, A. C., Meier, B., Gimenez-Abian, J. F., Diaz-Martinez, L. A., Green, J., Erickson, S. L., VanderWaal, K. E., Hsu, W.-S., Clarke, D. J. (2006). A mitotic topoisomerase II checkpoint in budding yeast is required for genome stability but acts independently of Pds1/securin.. Genes Dev. 20: 1162-1174 [Abstract] [Full Text]
Seth, R., Yang, C., Kaushal, V., Shah, S. V., Kaushal, G. P. (2005). p53-dependent Caspase-2 Activation in Mitochondrial Release of Apoptosis-inducing Factor and Its Role in Renal Tubular Epithelial Cell Injury. J. Biol. Chem. 280: 31230-31239 [Abstract] [Full Text]
Korfali, N., Ruchaud, S., Loegering, D., Bernard, D., Dingwall, C., Kaufmann, S. H., Earnshaw, W. C. (2004). Caspase-7 Gene Disruption Reveals an Involvement of the Enzyme during the Early Stages of Apoptosis. J. Biol. Chem. 279: 1030-1039 [Abstract] [Full Text]
Deming, P. B., Flores, K. G., Downes, C. S., Paules, R. S., Kaufmann, W. K. (2002). ATR Enforces the Topoisomerase II-dependent G2 Checkpoint through Inhibition of Plk1 Kinase. J. Biol. Chem. 277: 36832-36838 [Abstract] [Full Text]
Tomkiel, J. E., Alansari, H., Tang, N., Virgin, J. B., Yang, X., VandeVord, P., Karvonen, R. L., Granda, J. L., Kraut, M. J., Ensley, J. F., Fernandez-Madrid, F. (2002). Autoimmunity to the Mr 32,000 Subunit of Replication Protein A in Breast Cancer. Clin. Cancer Res. 8: 752-758 [Abstract] [Full Text]
Meng, X., Poon, R., Zhang, X., Cheah, A., Ding, Q., Hui, C.-c., Alman, B. (2001). Suppressor of Fused Negatively Regulates beta -Catenin Signaling. J. Biol. Chem. 276: 40113-40119 [Abstract] [Full Text]
Cao, G., Pei, W., Lan, J., Stetler, R. A., Luo, Y., Nagayama, T., Graham, S. H., Yin, X.-M., Simon, R. P., Chen, J. (2001). Caspase-Activated DNase/DNA Fragmentation Factor 40 Mediates Apoptotic DNA Fragmentation in Transient Cerebral Ischemia and in Neuronal Cultures. J. Neurosci. 21: 4678-4690 [Abstract] [Full Text]
Nagahara, Y., Ikekita, M., Shinomiya, T. (2000). Immunosuppressant FTY720 Induces Apoptosis by Direct Induction of Permeability Transition and Release of Cytochrome c from Mitochondria. J. Immunol. 165: 3250-3259 [Abstract] [Full Text]
van Gijn, R, Lendfers, R., Schellens, J., Bult, A, Beijnen, J. (2000). Dual topoisomerase I/II inhibitors. J Oncol Pharm Pract 6: 92-108 [Abstract]
Elizondo, G., Fernandez-Salguero, P., Sheikh, M. S., Kim, G.-Y., Fornace, A. J., Lee, K. S., Gonzalez, F. J. (2000). Altered Cell Cycle Control at the G2/M Phases in Aryl Hydrocarbon Receptor-Null Embryo Fibroblast. Mol. Pharmacol. 57: 1056-1063 [Abstract] [Full Text]
Gewies, A., Rokhlin, O. W., Cohen, M. B. (2000). Cytochrome c Is Involved in Fas-mediated Apoptosis of Prostatic Carcinoma Cell Lines. Cancer Res. 60: 2163-2168 [Abstract] [Full Text]
Lavoie, B. D., Tuffo, K. M., Oh, S., Koshland, D., Holm, C. (2000). Mitotic Chromosome Condensation Requires Brn1p, the Yeast Homologue of Barren. Mol. Biol. Cell 11: 1293-1304 [Abstract] [Full Text]
Rzepecki, R, Fisher, P. (2000). During both interphase and mitosis, DNA topoisomerase II interacts with DNA as well as RNA through the protein's C-terminal domain. J. Cell Sci. 113: 1635-1647 [Abstract]
Swanton, E., Bishop, N., Woodman, P. (1999). Human Rabaptin-5 Is Selectively Cleaved by Caspase-3 during Apoptosis. J. Biol. Chem. 274: 37583-37590 [Abstract] [Full Text]
Fernandez-Madrid, F., VandeVord, P. J., Yang, X., Karvonen, R. L., Simpson, P. M., Kraut, M. J., Granda, J. L., Tomkiel, J. E. (1999). Antinuclear Antibodies as Potential Markers of Lung Cancer. Clin. Cancer Res. 5: 1393-1400 [Abstract] [Full Text]
Samejima, K., Tone, S., Kottke, T. J., Enari, M., Sakahira, H., Cooke, C. A., Durrieu, F., Martins, L. M., Nagata, S., Kaufmann, S. H., Earnshaw, W. C. (1998). Transition from Caspase-dependent to Caspase-independent Mechanisms at the Onset of Apoptotic Execution. JCB 143: 225-239 [Abstract] [Full Text]
Dallaporta, B., Hirsch, T., Susin, S. A., Zamzami, N., Larochette, N., Brenner, C., Marzo, I., Kroemer, G. (1998). Potassium Leakage During the Apoptotic Degradation Phase. J. Immunol. 160: 5605-5615 [Abstract] [Full Text]
Meyer, K. N., Kjeldsen, E., Straub, T., Knudsen, B. R., Hickson, I. D., Kikuchi, A., Kreipe, H., Boege, F. (1997). Cell Cycle-coupled Relocation of Types I and II Topoisomerases and Modulation of Catalytic Enzyme Activities. JCB 136: 775-788 [Abstract] [Full Text]
Torok, T, Harvie, P D, Buratovich, M, Bryant, P J (1997). The product of proliferation disrupter is concentrated at centromeres and required for mitotic chromosome condensation and cell proliferation in Drosophila.. Genes Dev. 11: 213-225 [Abstract]
Nakajima, T., Morita, K., Ohi, N., Arai, T., Nozaki, N., Kikuchi, A., Osaka, F., Yamao, F., Oda, K. (1996). Degradation of Topoisomerase IIalpha during Adenovirus E1A-induced Apoptosis Is Mediated by the Activation of the Ubiquitin Proteolysis System. J. Biol. Chem. 271: 24842-24849 [Abstract] [Full Text]
Kimura, K., Nozaki, N., Enomoto, T., Tanaka, M., Kikuchi, A. (1996). Analysis of M Phase-specific Phosphorylation of DNA Topoisomerase II. J. Biol. Chem. 271: 21439-21445 [Abstract] [Full Text]
Kimura, K., Saijo, M., Tanaka, M., Enomoto, T. (1996). Phosphorylation-independent Stimulation of DNA Topoisomerase IIalpha Activity. J. Biol. Chem. 271: 10990-10995 [Abstract] [Full Text]
Meller, V., Fisher, P. (1995). Nuclear distribution of Drosophila DNA topoisomerase II is sensitive to both RNase and DNase. J. Cell Sci. 108: 1651-1657 [Abstract]
Morse-Gaudio, M, Risley, M. (1994). Topoisomerase II expression and VM-26 induction of DNA breaks during spermatogenesis in Xenopus laevis. J. Cell Sci. 107: 2887-2898 [Abstract]
Clarke, D., Johnson, R., Downes, C. (1993). Topoisomerase II inhibition prevents anaphase chromatid segregation in mammalian cells independently of the generation of DNA strand breaks. J. Cell Sci. 105: 563-569 [Abstract]
Buchenau, P, Saumweber, H, Arndt-Jovin, D. (1993). Consequences of topoisomerase II inhibition in early embryogenesis of Drosophila revealed by in vivo confocal laser scanning microscopy. J. Cell Sci. 104: 1175-1185 [Abstract]
Cardenas, M., Gasser, S. (1993). Regulation of topoisomerase II by phosphorylation: a role for casein kinase II. J. Cell Sci. 104: 219-225
Nigg, E.A., Krek, W., Peter, M. (1991). Vertebrate cdc2 Kinase: Its Regulation by Phosphorylation and Its Mitotic Targets. Cold Spring Harb Symp Quant Biol 56: 539-547 [Abstract]
Chen, D., Stetler, R. A., Cao, G., Pei, W., O'Horo, C., Yin, X.-M., Chen, J. (2000). Characterization of the Rat DNA Fragmentation Factor 35/Inhibitor of Caspase-activated DNase (Short Form). THE ENDOGENOUS INHIBITOR OF CASPASE-DEPENDENT DNA FRAGMENTATION IN NEURONAL APOPTOSIS. J. Biol. Chem. 275: 38508-38517 [Abstract] [Full Text]
Samejima, K., Tone, S., Earnshaw, W. C. (2001). CAD/DFF40 Nuclease Is Dispensable for High Molecular Weight DNA Cleavage and Stage I Chromatin Condensation in Apoptosis. J. Biol. Chem. 276: 45427-45432 [Abstract] [Full Text]