NAP-2 is part of multi-protein complexes in HeLa cells

Journal of Cellular Biochemistry
Pedro RodriguezMaria Zannis-Hadjopoulos

Abstract

We previously reported that a complex of nuclear proteins from HeLa cells, among them histone H1 and casein kinase 2 co-eluted from immobilized nucleosome assembly protein 2 (NAP-2)-Sepharose. Here, using HeLa cell nuclear extracts, we found NAP-2 migrates in a blue-native polyacrylamide gel with an apparent molecular weight of 300 kDa. HeLa cell NAP-2, labeled in vivo with radioactive orthophosphate, co-precipitated with at least two phosphoproteins, with an apparent mass of 100 and 175 kDa, respectively, as determined by SDS-PAGE. NAP-2 from total HeLa cell extract co-purified with other proteins through two sequential chromatographic steps: first, a positively charged resin, Q-Sepharose, was used, which purified NAP-2 more easily with other proteins that eluted as a single peak at 0.5 M NaCl. This fraction possessed both relaxing and supercoiling activities, and it was able to assemble regularly spaced nucleosomes onto naked DNA in an ATP-dependent manner. Second, a negatively charged resin (heparin) was used, which retained small amounts of NAP-2 (a very acidic polypeptide) and topoisomerase I. This fraction, although able to supercoil relaxed DNA, did so to a lesser extent than the Q-Sepharose fraction. The data suggest th...Continue Reading

References

Nov 1, 1978·Cell·A WorcelM L Wong
May 1, 1975·Proceedings of the National Academy of Sciences of the United States of America·J E GermondP Chambon
May 25, 1979·Science·A PrunellF H Crick
Aug 1, 1989·Biochemistry and Cell Biology = Biochimie Et Biologie Cellulaire·E B KmiecA D Cole
Nov 1, 1987·Molecular and Cellular Biology·R B MorelandL M Hereford
Jun 5, 1986·Journal of Molecular Biology·I Ruberti, A Worcel
Jan 2, 1987·European Journal of Biochemistry·Y IshimiF Hanaoka
Sep 25, 1981·Nucleic Acids Research·V JacksonR Chalkley
Nov 15, 1993·Proceedings of the National Academy of Sciences of the United States of America·G Arents, E N Moudrianakis
Jan 1, 1996·Annual Review of Biochemistry·J C Wang
Dec 31, 1997·Molecular and Cellular Biology·H Kurumizaka, A P Wolffe
Apr 16, 1998·The Journal of Biological Chemistry·G A McQuibbanP N Lewis
Mar 31, 1998·Current Opinion in Structural Biology·K Luger, T J Richmond
Apr 5, 2000·The Journal of Biological Chemistry·C RegnardB Eddé
Apr 15, 2000·Journal of Molecular Biology·P RodriguezM Zannis-Hadjopoulos

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Citations

Apr 19, 2005·The International Journal of Biochemistry & Cell Biology·Helmut Grasberger, Graeme I Bell
Nov 17, 2005·Plant Methods·Holger EubelA Harvey Millar
Oct 3, 2006·Journal of Biochemistry·Guillermo CoveloManuel Freire
Feb 24, 2007·FASEB Journal : Official Publication of the Federation of American Societies for Experimental Biology·Jordanka ZlatanovaMiroslav Tomschik
Jul 8, 2018·Drug Metabolism and Disposition : the Biological Fate of Chemicals·Duan LiuRichard M Weinshilboum
Jun 19, 2008·Molecular and Cellular Biochemistry·Birgitte B Olsen, Barbara Guerra
Sep 25, 2007·Nano Letters·Bhranti S ShahJeremy J Mao

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