CGI-55 interacts with nuclear proteins and co-localizes to p80-coilin positive-coiled bodies in the nucleus
The human protein CGI-55 has been described as a chromo-helicase-DNA-binding domain protein (CHD)-3 interacting protein and was also found to interact with the 3′-region of the plasminogen activator inhibitor (PAI)-1 mRNA. Here, we used CGI-55 as a “bait” in a yeast two-hybrid screen and identified eight interacting proteins: Dax, Topoisomerase I binding RS (Topors), HPC2, UBA2, TDG, and protein inhibitor of activated STAT (signal transducer and activator of transcription) (PIAS)-1,-3, and-y. These proteins are either structurally or functionally associated with promyelocytic leukemia nuclear bodies (PML-NBs), protein sumoylation, or the regulation of transcription. The interactions of CGI-55 with Daxx, Topors, PIASy, and UBA2 were confirmed by in vivo colocalization experiments in HeLa cells, by using green (GFP) and red fluorescence fusion proteins. A mapping study of the CGI-55 binding site for these proteins revealed three distinct patterns of interaction. The fact that CGI-55-GFP has been localized in cytoplasm and nucleus in a dotted manner, and its interaction with proteins associated with PML-NBs, suggested that CGI-55 might be associated with nuclear bodies. Although Daxx and Topors co-localized with promyelocytic leukemia protein (PML), CGI-55 itself as well as PIASy and UBA2 showed only little co-localization with PML. However, we observed that CGI-55 localizes to the nucleolus and co-localizes with p80-coilin positive nuclear-coiled bodies.
Index EntriesPML-NBs coiled-bodies Cajal bodies protein-protein interaction two-hybrid domain mapping immunolocalization p80-coilin
Kobarg, J., Schnittger, S., Fonatsch, C., et al. (1997) Characterization, mapping and partial cDNA sequence of the 57-kDa intracellular Ki-1 antigen. Exp. Clin. Immunogenet.
, 273–280.PubMedGoogle Scholar
Lemos, T. A., Passos, D. O., Nery, F. C., and Kobarg, J. (2003) Characterization of a new family of proteins that interact with the C-terminal region of the chromatin-remodeling factor CHD-3. FEBS Lett.
, 14–20.PubMedCrossRefGoogle Scholar
Heaton, J. H., Dlakic, W. M., Dlakic, M., and Gelehrter, T. D. (2001) Identification and cDNA cloning of a novel RNA-binding protein that interacts with the cyclic nucleotide-responsive sequence in the type-1 plasminogen activator inhibitor mRNA. J. Biol. Chem.
, 3341–3347.PubMedCrossRefGoogle Scholar
Huang, L., Grammatikakis, N., Yoneda, M., Banerjee, S. D., and Toole, B. P. (2000) Molecular characterization of a novel intracellular hyaluronan-binding protein. J. Biol. Chem.
, 29,829–29,839.Google Scholar
Nery, F. C., Passos, D. O., Garcia, V. S., and Kobarg, J. (2004) Ki-1/57 interacts with RACK1 and is a substrate for PMA activated PKC. J. Biol. Chem.
, 11,444–11,455.CrossRefGoogle Scholar
Ozaki, T., Watanabe, K.-I., Nakagawa, T., Miyazaki, K., Takahashi, M., and Nakagawara, A. (2003) Function of p73, not of p53, is inhibited by the physical interaction with RACK1 and its inhibitory effect is counteracted by pRB. Oncogene
, 3231–3242.PubMedCrossRefGoogle Scholar
Matera, A. G. (1999) Nuclear bodies: multifaceted subdomains of the interchromatin space. Trends Cell Biol.
, 302–309.PubMedCrossRefGoogle Scholar
Andrade, L. E. C., Tan, E. M., and Chan, E. K. L. (1993) Immunocytochemical analysis of the coiled body in the cell cycle and during cell proliferation. Proc. Natl. Acad. Sci. U.S.A.
, 1947–1951.CrossRefGoogle Scholar
Ogg, S. C., and Lamond, A. I. (2002) Cajal bodies and coilin-moving towards function. J. Cell Biol.
, 17–21.CrossRefGoogle Scholar
Zhong, S., Salomoni, P., and Pandolfi, P. (2000) The transcriptional role of PML and the nuclear body. Nat. Cell Biol.
, E85-E90.PubMedCrossRefGoogle Scholar
Rasheed, Z. A., Saleem, A., Ravee, Y., Pandolfi, P. P., and Rubin, E. H. (2002) The topoisomerase I-binding RING protein, topors, is associated with promyelocytic leukemia nuclear bodies. Exp. Cell. Res.
, 152–160.PubMedCrossRefGoogle Scholar
Salomoni, P. and Pandolfi, P. P. (2002) The role of PML in tumor suppression. Cell
, 165–170.PubMedCrossRefGoogle Scholar
Everett, R. D., Lomonte, P., Sternsdorf, T., van Driel, R., and Orr, A. (1999) Cell cycle regulation of PML modification and ND10 composition. J. Cell Sci.
, 4581–4588.PubMedGoogle Scholar
Zhong, S., Müller, S., Ronchetti, S., Freemont, P. S., Dejean, A., and Pandolfi, P. P. (2000) Role of SUMO-1-modified PML in nuclear body formation. Blood
, 2748–2752.PubMedGoogle Scholar
Borden, K. L. (2002) Pondering the promyelocytic leukemia protein (PML) puzzle: possible functions from PML nuclear bodies. Mol. Cell. Biol.
, 5259–5269.PubMedCrossRefGoogle Scholar
Sterndorf, T., Jensen, K., and Will, H. (1997) Evidence for covalent modification of the nuclear dot-associated proteins PML and SP100 by PIC1/SUMO1. J. Cell Biol.
, 1621–1634.CrossRefGoogle Scholar
Ishov, A. M., Sotnikov, A. G., Negorev, D., et al. (1999) PML is critical for ND10 formation and recruits the PML-interacting protein daxx to this nuclear structure when modified by SUMO-1. J. Cell Biol.
, 221–234.PubMedCrossRefGoogle Scholar
Fields, S. and Song, O. (1989) A novel genetic system to detect protein-protein interactions. Nature
, 245–246.PubMedCrossRefGoogle Scholar
Vojtek, A. B. and Hollenberg, S. M. (1995) Ras-Raf interaction: two-hybrid analysis. Methods Enzymol.
, 331–342.PubMedCrossRefGoogle Scholar
Moraes, K. C., Quaresma, A. J., Maehnss, K., and Kobarg, J. (2003) Identification and characterization of proteins that selectively interact with isoforms of the mRNA binding protein AUF1 (hnRNP D). Biol. Chem.
, 35–37.CrossRefGoogle Scholar
Schmidt, D. and Müller, S. (2002) Members of the PIAS family act as SUMO ligases for c-Jun and p53 and repress p53 activity. Proc. Natl. Acad. Sci. U.S.A.
, 2872–2877.PubMedCrossRefGoogle Scholar
Kotaja, N., Karvonen, U., Janne, O. A., and Palvimo, J. J. (2002) PIAS proteins modulate transcription factors by functioning as SUMO-1 ligases. Mol Cell Biol.
, 5222–5234.PubMedCrossRefGoogle Scholar
Müller, S., Matunis, M. J., and Dejean, A. (1998) Conjugation with the ubiquitin-related modifier SUMO-1 regulates the partitioning of PML within the nucleus. EMBO J.
, 61–70.PubMedCrossRefGoogle Scholar
Valdez, B. C., Henning, D., Perlaky, L., Busch, R. K., and Busch, H. (1997) Cloning and characterization of Gu/RH-II binding protein. Biochem. Biophys. Res. Commun.
, 335–340.PubMedCrossRefGoogle Scholar
Miyauchi, Y., Yogosawa, S., Honda, R., Nishida, T., and Yasuda, H. (2002) Sumoylation of Mdm2 by protein inhibitor of activated STAT (PIAS) and RanBP2 enzymes. J. Biol. Chem.
, 50,131–50,136.CrossRefGoogle Scholar
Haluska, P., Jr., Saleem, A., Rasheed, Z., et al. (1999) Interaction between human topoisomerase I and a novel RING-finger/arginine-serine protein. Nucleic Acids Res.
, 2538–2544.PubMedCrossRefGoogle Scholar
Zhou, R., Wen, H., and Ao, S. Z. (1999) Identification of a novel gene encoding a p53-associated protein. Gene
, 93–101.PubMedCrossRefGoogle Scholar
Rechsteiner, M., Rogers, S. W. (1996) PEST sequences and regulation by proteolysis. Trends Biochem. Sci.
, 267–271.PubMedCrossRefGoogle Scholar
Torii, S., Egan, D. A., Evans, R. A., and Reed, J. C. (1999) Human Daxx regulates Fas-induced apoptosis from nuclear PML oncogenic domains (PODs). EMBO J.
, 6037–6049.PubMedCrossRefGoogle Scholar
Li, R., Pei, H., Watson, D. K., and Papas, T. S. (2000) EAP1/Daxx interacts with ETS1 and represses transcriptional activation of ETS1 target genes. Oncogene
, 745–753.PubMedCrossRefGoogle Scholar
Ko, Y. G., Kang, Y. S., Park, H., et al. (2001) Apoptosis signal-regulating kinase 1 controls the proapoptotic function of death-associated protein (Daxx) in the cytoplasm. J. Biol. Chem.
, 39,103–39,106.Google Scholar
Lin, D. Y., Lai, M. Z., Ann, D. K., and Shih, H. M. (2003) Promyelocytic leukemia protein (PML) functions as a glucocorticoid receptor co-activator by Sequestering Daxx to the PML oncogenic domains (PODs) to enhance its transactivation potential. J. Biol. Chem.
, 15,958–15,965.Google Scholar
Shih, H. P., Hales, K. G., Pringle, J. R., and Peifer, M. (2002) Identification of septin-interacting proteins and characterization of the Smt3/SUMO-conjugation system in Drosophila
. J. Cell Sci.
, 1259–1271.PubMedGoogle Scholar
Chu, D., Kakazu, N., Gorrin-Rivas, M. J., et al. (2001) Cloning and characterization of LUN, a novel ring finger protein that is highly expressed in lung and specifically binds to a palindromic sequence. J. Biol. Chem.
, 14,004–14,013.Google Scholar
Yang, X., Khosravi-Far, R., Chang, H. Y., and Baltimore, D. (1997) Daxx, a novel Fas-binding protein that activates JNK and apoptosis. Cell
, 1067–1076.PubMedCrossRefGoogle Scholar
Emelyanov, A. V., Kovac, C. R., Sepulveda, M. A., and Birshtein, B. K. (2002) The interaction of Pax5 (BSAP) with Daxx can result in transcriptional activation in B cells. J. Biol. Chem.
, 11,156–11,164.CrossRefGoogle Scholar
Pluta, A. F., Earnshaw, W. C., and Goldberg, I. G. (1998) Interphase-specific association of intrinsic centromer protein CENP-C with Daxx, a death domain-binding protein implicated in Fas-mediated cell death. J. Cell. Sci.
, 2029–2041.PubMedGoogle Scholar
Satijn, D. P., Olson, D. J., van der Vlag, J., et al. (1997) Interference with the expression of a novel human polycomb protein, hPc2, results in cellular transformation and apoptosis. Mol. Cell. Biol.
, 6076–6086.PubMedGoogle Scholar
Liu, B., Liao, J., Rao, X., et al. (1998) Inhibition of Stat1-mediated gene activation by PIAS1. Proc. Natl. Acad. Sci. U.S.A.
, 10,626–10,631.Google Scholar
Kahyo, T., Nishida, T., and Yasuda, H. (2001) Involvement of PIAS1 in the sumoylation of tumor suppressor p53. Mol. Cell.
, 713–718.PubMedCrossRefGoogle Scholar
Jackson, P. K. (2001) A new RING for SUMO: wrestling transcriptional responses into nuclear bodieswith PIAS family E3 SUMO ligases. Genes Dev.
, 3053–3058.PubMedCrossRefGoogle Scholar
Desterro, J. M., Rodríguez, M. S., Kemp, G. D., and Hay, R. T. (1999) Identification of the enzyme required for activation of the small ubiquitin-like protein SUMO-1. J. Biol. Chem.
, 10,618–10,624.CrossRefGoogle Scholar
Gong, L., Li, B., Millas, S., and Yeh, E. T. (1999) Molecular cloning and characterization of human AOS1 and UBA2, components of the sentrin-activating enzyme complex. FEBS Lett.
, 185–189.PubMedCrossRefGoogle Scholar
Okuma, T., Honda, R., Ichikawa, G., Tsumagari, N., and Yasuda, H. (1999) In vitro SUMO-1 modification requires two enzymatic steps, E1 and E2. Biochem. Biophys. Res. Commun.
, 693–698.PubMedCrossRefGoogle Scholar
Rodriguez, M. S., Desterro, J. M., Lían, S., Midgley, C. A., Lane, D. P., and Hay, R. T. (1999) SUMO-1 modification activates the transcriptional response of p53. EMBO J.
, 6455–6461.PubMedCrossRefGoogle Scholar
Neddermann, P., Gallinari, P., Lettieri, T., et al. (1996) Cloning and expression of human G/T mismatch-specific thymine-DNA glycosylase. J. Biol. Chem.
, 12,767–12,774.Google Scholar
Lindahl, T. (1982) DNA repair enzymes. Annu. Rev. Biochem.
, 61–87.PubMedCrossRefGoogle Scholar
Hardeland, U., Steinacher, R., Jiricny, J., and Schär, P. (2002) Modification of the human tymine-DNA-glycosylase by ubiquitin-like proteins facilitates enzymatic turnover. EMBO J.
, 1456–1464.PubMedCrossRefGoogle Scholar
Takahashi, H., Hatakeyama, S., Saitoh, H., and Nakayama, K. I. (2005) Noncovalent SUMO-1 binding of thymine DNA glycosylase (TDG) is required for its SUMO-1 modification and colocalization with the promyelocytic leukemia protein (PML). J. Biol. Chem.
, 5611–5621.PubMedCrossRefGoogle Scholar
Boddy, M. N., Howe, K., Etkin, L. D., Solomon, E., and Freemont, P. S. (1996) PIC 1, a novel ubiquitin-like protein which interacts with the PML component of a multiprotein complex that is disrupted in acute promyelocytic leukaemia. Oncogene
, 971–982.PubMedGoogle Scholar
Long, J., Matsura, I., He, D., Wang, G., Shuai, K., and Liu, F. (2003) Repression of SMAD transcriptional activity by PIASy, an inhibitor of activated STAT. Proc. Natl. Acad. Sci. U.S.A.
, 9791–9796.PubMedCrossRefGoogle Scholar