The Journal of Cell Biology, Vol 126, 1331-1340, Copyright © 1994 by The Rockefeller University Press

ARTICLES

An RNase-sensitive particle containing Drosophila melanogaster DNA topoisomerase II

VH Meller, M McConnell and PA Fisher
Department of Pharmacological Sciences, University Medical Center, State University of New York at Stony Brook 11794-8651.

Most DNA topoisomerase II (topo II) in cell-free extracts of 0-2-h old Drosophila embryos appears to be nonnuclear and remains in the supernatant after low-speed centrifugation (10,000 g). Virtually all of this apparently soluble topo II is particulate with a sedimentation coefficient of 67 S. Similar topo II-containing particles were detected in Drosophila Kc tissue culture cells, 16-19-h old embryos and extracts of progesterone-matured oocytes from Xenopus. Drosophila topo II- containing particles were insensitive to EDTA, Triton X-100 and DNase I, but could be disrupted by incubation with 0.3 M NaCl or RNase A. After either disruptive treatment, topo II sedimented at 9 S. topo II- containing particles were also sensitive to micrococcal nuclease. Results of chemical cross-linking corroborated those obtained by centrifugation. Immunoblot analyses demonstrated that topo II- containing particles lacked significant amounts of lamin, nuclear pore complex protein gp210, proliferating cell nuclear antigen, RNA polymerase II subunits, histones, coilin, and nucleolin. Northern blot analyses demonstrated that topo II-containing particles lacked U RNA. Thus, current data support the notion that nonnuclear Drosophila topo II-containing particles are composed largely of topo II and an unknown RNA molecule(s).
CiteULike    Complore    Connotea    Del.icio.us    Digg    Facebook    Reddit    Technorati    Twitter    What's this?

This article has been cited by other articles:

• 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]  
• Rzepecki, R, Bogachev, S., Kokoza, E, Stuurman, N, Fisher, P. (1998). In vivo association of lamins with nucleic acids in Drosophila melanogaster. J. Cell Sci. 111: 121-129 [Abstract]  
• 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]  
• Barthelmes, H. U., Grue, P., Feineis, S., Straub, T., Boege, F. (2000). Active DNA Topoisomerase IIalpha Is a Component of the Salt-stable Centrosome Core. J. Biol. Chem. 275: 38823-38830 [Abstract] [Full Text]  

Home | Help | Feedback | Subscriptions | Archive | Search | Table of Contents