Journal of Cell Science, Vol 114, Issue 1 59-68, Copyright © 2001 by Company of Biologists

JOURNAL ARTICLES

Evidence for separate ND10-binding and homo-oligomerization domains of Sp100

D Negorev, AM Ishov and GG Maul
The Wistar Institute, Philadelphia, PA, USA.

Nuclear domains called ND10 or PML nuclear bodies consist of an aggregation of several proteins, most notably PML and Sp100. PML is essential in the nucleation and formation of ND10 as well as in the recruitment of other ND10-associated proteins such as Daxx, pRb, BLM and Sp100. In cells induced to overexpress Sp100, ND10 binding of Sp100 was saturable and excess Sp100 formed new aggregation sites devoid of other ND10-associated proteins, suggesting that homo-oligomerization is the basis for aggregation. To determine whether Sp100 binds to ND10 through hetero- or oligomerization, Sp100 deletion variants fused with GFP were transfected into cells with and without endogenous Sp100, and the localization of the GFP-labeled fragments was determined relative to ND10. Amino acids 29-152 were sufficient for deposition of the GFP-labeled fragments at ND10 in the absence of endogenous Sp100 (heterologous binding) and for self-aggregation (formation of new Sp100 deposits). None of the shorter fragments was deposited at ND10 or self-aggregated. The 29-152 amino acid fragment and some larger fragments, but not the full-size Sp100, induced elongation of ND10, which at their ends contain only Sp100, probably due to self-aggregation. By fusing a peptide consisting of the p53-binding domain from hMDM2 to the Sp100(29-152) fragment, this self-aggregation could be blocked while retaining the limited ND10 binding capacity, indicating that the Sp100 self-aggregation domain and the ND10 binding domain are separate entities. This fusion peptide was used to demonstrate the potential of ND10 to recruit p53 as a protein not usually present at this site. Such deposited p53 was protected from turnover. The capacity of ND10 to recruit Sp100 may serve primarily to reduce its availability.
CiteULike    Complore    Connotea    Del.icio.us    Digg    Reddit    Technorati    Twitter    What's this?

This article has been cited by other articles:

				G. Cho, Y. Lim, and J. A. Golden SUMO Interaction Motifs in Sizn1 Are Required for Promyelocytic Leukemia Protein Nuclear Body Localization and for Transcriptional Activation J. Biol. Chem., July 17, 2009; 284(29): 19592 - 19600. [Abstract] [Full Text] [PDF]

				D. G. Negorev, O. V. Vladimirova, and G. G. Maul Differential Functions of Interferon-Upregulated Sp100 Isoforms: Herpes Simplex Virus Type 1 Promoter-Based Immediate-Early Gene Suppression and PML Protection from ICP0-Mediated Degradation J. Virol., May 15, 2009; 83(10): 5168 - 5180. [Abstract] [Full Text] [PDF]

				S. Weidtkamp-Peters, T. Lenser, D. Negorev, N. Gerstner, T. G. Hofmann, G. Schwanitz, C. Hoischen, G. Maul, P. Dittrich, and P. Hemmerich Dynamics of component exchange at PML nuclear bodies J. Cell Sci., August 15, 2008; 121(16): 2731 - 2743. [Abstract] [Full Text] [PDF]

				D. G. Negorev, O. V. Vladimirova, A. Ivanov, F. Rauscher III, and G. G. Maul Differential role of sp100 isoforms in interferon-mediated repression of herpes simplex virus type 1 immediate-early protein expression. J. Virol., August 1, 2006; 80(16): 8019 - 8029. [Abstract] [Full Text] [PDF]

				M. Breuleux, F. Schoumacher, D. Rehn, W. Kung, H. Mueller, and U. Eppenberger Heregulins Implicated in Cellular Functions Other Than Receptor Activation Mol. Cancer Res., January 1, 2006; 4(1): 27 - 37. [Abstract] [Full Text] [PDF]

				W.-Q. Jiang, Z.-H. Zhong, J. D. Henson, A. A. Neumann, A. C.-M. Chang, and R. R. Reddel Suppression of Alternative Lengthening of Telomeres by Sp100-Mediated Sequestration of the MRE11/RAD50/NBS1 Complex Mol. Cell. Biol., April 1, 2005; 25(7): 2708 - 2721. [Abstract] [Full Text] [PDF]

				M. A. Suico, H. Yoshida, Y. Seki, T. Uchikawa, Z. Lu, T. Shuto, K. Matsuzaki, M. Nakao, J.-D. Li, and H. Kai Myeloid Elf-1-like Factor, an ETS Transcription Factor, Up-regulates Lysozyme Transcription in Epithelial Cells through Interaction with Promyelocytic Leukemia Protein J. Biol. Chem., April 30, 2004; 279(18): 19091 - 19098. [Abstract] [Full Text] [PDF]

				J. H. Yu, A. Nakajima, H. Nakajima, L. R. Diller, K. D. Bloch, and D. B. Bloch Restoration of Promyelocytic Leukemia Protein-Nuclear Bodies in Neuroblastoma Cells Enhances Retinoic Acid Responsiveness Cancer Res., February 1, 2004; 64(3): 928 - 933. [Abstract] [Full Text] [PDF]

				J. J. Partridge, J. O. Lopreiato Jr., M. Latterich, and F. E. Indig DNA Damage Modulates Nucleolar Interaction of the Werner Protein with the AAA ATPase p97/VCP Mol. Biol. Cell, October 1, 2003; 14(10): 4221 - 4229. [Abstract] [Full Text] [PDF]

				C. Ramsey, A. Bukrinsky, and L. Peltonen Systematic mutagenesis of the functional domains of AIRE reveals their role in intracellular targeting Hum. Mol. Genet., December 15, 2002; 11(26): 3299 - 3308. [Abstract] [Full Text] [PDF]

				C. Wasylyk, S. E. Schlumberger, P. Criqui-Filipe, and B. Wasylyk Sp100 Interacts with ETS-1 and Stimulates Its Transcriptional Activity Mol. Cell. Biol., April 15, 2002; 22(8): 2687 - 2702. [Abstract] [Full Text] [PDF]

				J.-S. Seeler, A. Marchio, R. Losson, J. M. P. Desterro, R. T. Hay, P. Chambon, and A. Dejean Common Properties of Nuclear Body Protein SP100 and TIF1{alpha} Chromatin Factor: Role of SUMO Modification Mol. Cell. Biol., May 15, 2001; 21(10): 3314 - 3324. [Abstract] [Full Text]