Abstract
Hub proteins have central roles in regulating cellular processes. By targeting a single cellular hub, a viral oncogene may gain control over an entire module in the cellular interaction network that is potentially comprised of hundreds of proteins. The adenovirus E1A oncoprotein is a viral hub that interacts with many cellular hub proteins by short linear motifs/molecular recognition features (MoRFs). These interactions transform the architecture of the cellular protein interaction network and virtually reprogram the cell. To identify additional MoRFs within E1A, we screened portions of E1A for their ability to activate yeast pseudohyphal growth or differentiation. This identified a novel functional region within E1A conserved region 2 comprised of the sequence EVIDLT. This MoRF is necessary and sufficient to bind the N-terminal region of the SUMO conjugase UBC9, which also interacts with SUMO noncovalently and is involved in polySUMOylation. Our results suggest that E1A interferes with polySUMOylation, but not with monoSUMOylation. These data provide the first insight into the consequences of the interaction of E1A with UBC9, which was initially described in 1996. We further demonstrate that polySUMOylation regulates pseudohyphal growth and promyelocytic leukemia body reorganization by E1A. In conclusion, the interaction of the E1A oncogene with UBC9 mimics the normal binding between SUMO and UBC9 and represents a novel mechanism to modulate polySUMOylation.
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References
Ansieau S, Leutz A . (2002). The conserved Mynd domain of BS69 binds cellular and oncoviral proteins through a common PXLXP motif. J Biol Chem 277: 4906–4910.
Avvakumov N, Kajon AE, Hoeben RC, Mymryk JS . (2004). Comprehensive sequence analysis of the E1A proteins of human and simian adenoviruses. Virology 329: 477–492.
Avvakumov N, Wheeler R, D'Halluin JC, Mymryk JS . (2002). Comparative sequence analysis of the largest E1A proteins of human and simian adenoviruses. J Virol 76: 7968–7975.
Bayley ST, Mymryk JS . (1994). Adenovirus E1A proteins and transformation. Int J Oncol 5: 425–444.
Berk AJ . (2005). Recent lessons in gene expression, cell cycle control, and cell biology from adenovirus. Oncogene 24: 7673–7685.
Bernardi R, Pandolfi PP . (2007). Structure, dynamics and functions of promyelocytic leukaemia nuclear bodies. Nat Rev Mol Cell Biol 8: 1006–1016.
Bylebyl GR, Belichenko I, Johnson ES . (2003). The SUMO isopeptidase Ulp2 prevents accumulation of SUMO chains in yeast. J Biol Chem 278: 44113–44120.
Capili AD, Lima CD . (2007). Structure and analysis of a complex between SUMO and Ubc9 illustrates features of a conserved E2-Ubl interaction. J Mol Biol 369: 608–618.
Chinnadurai G . (2002). CtBP, an unconventional transcriptional corepressor in development and oncogenesis. Mol Cell 9: 213–224.
Dellaire G, Bazett-Jones DP . (2004). PML nuclear bodies: dynamic sensors of DNA damage and cellular stress. Bioessays 26: 963–977.
Dunnebier T, Bermejo JL, Haas S, Fischer HP, Pierl CB, Justenhoven C et al. (2009). Common variants in the UBC9 gene encoding the SUMO-conjugating enzyme are associated with breast tumor grade. Int J Cancer 125: 596–602.
Eskiw CH, Dellaire G, Mymryk JS, Bazett-Jones DP . (2003). Size, position and dynamic behavior of PML nuclear bodies following cell stress as a paradigm for supramolecular trafficking and assembly. J Cell Sci 116: 4455–4466.
Ferrari R, Pellegrini M, Horwitz GA, Xie W, Berk AJ, Kurdistani SK . (2008). Epigenetic reprogramming by adenovirus E1A. Science 321: 1086–1088.
Fraser HB, Hirsh AE, Steinmetz LM, Scharfe C, Feldman MW . (2002). Evolutionary rate in the protein interaction network. Science 296: 750–752.
Frisch SM, Mymryk JS . (2002). Adenovirus-5 E1A: paradox and paradigm. Nat Rev Mol Cell Biol 3: 441–452.
Galanty Y, Belotserkovskaya R, Coates J, Polo S, Miller KM, Jackson SP . (2009). Mammalian SUMO E3-ligases PIAS1 and PIAS4 promote responses to DNA double-strand breaks. Nature 462: 935–939.
Gandhi TK, Zhong J, Mathivanan S, Karthick L, Chandrika KN, Mohan SS et al. (2006). Analysis of the human protein interactome and comparison with yeast, worm and fly interaction datasets. Nat Genet 38: 285–293.
Geiss-Friedlander R, Melchior F . (2007). Concepts in sumoylation: a decade on. Nat Rev Mol Cell Biol 8: 947–956.
Han JD, Bertin N, Hao T, Goldberg DS, Berriz GF, Zhang LV et al. (2004). Evidence for dynamically organized modularity in the yeast protein-protein interaction network. Nature 430: 88–93.
Hateboer G, Hijmans EM, Nooij JB, Schlenker S, Jentsch S, Bernards R . (1996). mUBC9, a novel adenovirus E1A-interacting protein that complements a yeast cell cycle defect. J Biol Chem 271: 25906–25911.
Jeong H, Mason SP, Barabasi AL, Oltvai ZN . (2001). Lethality and centrality in protein networks. Nature 411: 41–42.
Kerscher O, Felberbaum R, Hochstrasser M . (2006). Modification of proteins by ubiquitin and ubiquitin-like proteins. Annu Rev Cell Dev Biol 22: 159–180.
Knipscheer P, Flotho A, Klug H, Olsen JV, van Dijk WJ, Fish A et al. (2008). Ubc9 sumoylation regulates SUMO target discrimination. Mol Cell 31: 371–382.
Knipscheer P, van Dijk WJ, Olsen JV, Mann M, Sixma TK . (2007). Noncovalent interaction between Ubc9 and SUMO promotes SUMO chain formation. EMBO J 26: 2797–2807.
Lallemand-Breitenbach V, Jeanne M, Benhenda S, Nasr R, Lei M, Peres L et al. (2008). Arsenic degrades PML or PML-RAR alpha through a SUMO-triggered RNF4/ubiquitin-mediated pathway. Nat Cell Biol 10: 547–555.
Lin D, Tatham MH, Yu B, Kim S, Hay RT, Chen Y . (2002). Identification of a substrate recognition site on Ubc9. J Biol Chem 277: 21740–21748.
Lowe SW, Jacks T, Housman DE, Ruley HE . (1994). Abrogation of oncogene-associated apoptosis allows transformation of p53-deficient cells. Proc Natl Acad Sci USA 91: 2026–2030.
Maslov S, Sneppen K . (2002). Specificity and stability in topology of protein networks. Science 296: 910–913.
Mo YY, Moschos SJ . (2005). Targeting Ubc9 for cancer therapy. Expert Opin Ther Targets 9: 1203–1216.
Mo YY, Yu Y, Ee PL, Beck WT . (2004). Overexpression of a dominant-negative mutant Ubc9 is associated with increased sensitivity to anticancer drugs. Cancer Res 64: 2793–2798.
Morris EJ, Dyson NJ . (2001). Retinoblastoma protein partners. Adv Cancer Res 82: 1–54.
Morris JR, Boutell C, Keppler M, Densham R, Weekes D, Alamshah A et al. (2009). The SUMO modification pathway is involved in the BRCA1 response to genotoxic stress. Nature 462: 886–890.
Mullen JR, Brill SJ . (2008). Activation of the Slx5-Slx8 ubiquitin ligase by poly-small ubiquitin-like modifier conjugates. J Biol Chem 283: 19912–19921.
Mymryk JS, Smith MM . (1997). Influence of the adenovirus 5 E1A oncogene on chromatin remodelling. Biochem Cell Biol 75: 95–102.
O'Connor MJ, Zimmermann H, Nielsen S, Bernard HU, Kouzarides T . (1999). Characterization of an E1A-CBP interaction defines a novel transcriptional adapter motif (TRAM) in CBP/p300. J Virol 73: 3574–3581.
Pelka P, Ablack JN, Fonseca GJ, Yousef AF, Mymryk JS . (2008). Intrinsic structural disorder in adenovirus E1A: a viral molecular hub linking multiple diverse processes. J Virol 82: 7252–7263.
Prudden J, Pebernard S, Raffa G, Slavin DA, Perry JJ, Tainer JA et al. (2007). SUMO-targeted ubiquitin ligases in genome stability. EMBO J 26: 4089–4101.
Prudden J, Perry JJ, Arvai AS, Tainer JA, Boddy MN . (2009). Molecular mimicry of SUMO promotes DNA repair. Nat Struct Mol Biol 16: 509–516.
Rual JF, Venkatesan K, Hao T, Hirozane-Kishikawa T, Dricot A, Li N et al. (2005). Towards a proteome-scale map of the human protein-protein interaction network. Nature 437: 1173–1178.
Rupp S, Summers E, Lo H-J, Madhani H, Fink G . (1999). MAP kinase and cAMP filamentation signaling pathways converge on the unusually large promoter of the yeast FLO11 gene. EMBO J 18: 1257–1269.
Saitoh H, Hinchey J . (2000). Functional heterogeneity of small ubiquitin-related protein modifiers SUMO-1 versus SUMO-2/3. J Biol Chem 275: 6252–6258.
Sampson DA, Wang M, Matunis MJ . (2001). The small ubiquitin-like modifier-1 (SUMO-1) consensus sequence mediates Ubc9 binding and is essential for SUMO-1 modification. J Biol Chem 276: 21664–21669.
Schmidt D, Muller S . (2002). Members of the PIAS family act as SUMO ligases for c-Jun and p53 and repress p53 activity. Proc Natl Acad Sci USA 99: 2872–2877.
Schwienhorst I, Johnson ES, Dohmen RJ . (2000). SUMO conjugation and deconjugation. Mol Gen Genet 263: 771–786.
Sekiyama N, Arita K, Ikeda Y, Hashiguchi K, Ariyoshi M, Tochio H et al. (2009). Structural basis for regulation of poly-SUMO chain by a SUMO-like domain of Nip45. Proteins 78: 1491–1502.
Song J, Durrin LK, Wilkinson TA, Krontiris TG, Chen Y . (2004). Identification of a SUMO-binding motif that recognizes SUMO-modified proteins. Proc Natl Acad Sci USA 101: 14373–14378.
Song J, Zhang Z, Hu W, Chen Y . (2005). Small ubiquitin-like modifier (SUMO) recognition of a SUMO binding motif: a reversal of the bound orientation. J Biol Chem 280: 40122–40129.
Stelzl U, Worm U, Lalowski M, Haenig C, Brembeck FH, Goehler H et al. (2005). A human protein-protein interaction network: a resource for annotating the proteome. Cell 122: 957–968.
Sun H, Leverson JD, Hunter T . (2007). Conserved function of RNF4 family proteins in eukaryotes: targeting a ubiquitin ligase to SUMOylated proteins. EMBO J 26: 4102–4112.
Tatham MH, Geoffroy MC, Shen L, Plechanovova A, Hattersley N, Jaffray EG et al. (2008). RNF4 is a poly-SUMO-specific E3 ubiquitin ligase required for arsenic-induced PML degradation. Nat Cell Biol 10: 538–546.
Trentin JL, Yabe Y, Taylor G . (1962). The quest for human cancer viruses. Science 137: 835–841.
Uzunova K, Gottsche K, Miteva M, Weisshaar SR, Glanemann C, Schnellhardt M et al. (2007). Ubiquitin-dependent proteolytic control of SUMO conjugates. J Biol Chem 282: 34167–34175.
Wang J, Hu W, Cai S, Lee B, Song J, Chen Y . (2007). The intrinsic affinity between E2 and the Cys domain of E1 in ubiquitin-like modifications. Mol Cell 27: 228–237.
Weisshaar SR, Keusekotten K, Krause A, Horst C, Springer HM, Gottsche K et al. (2008). Arsenic trioxide stimulates SUMO-2/3 modification leading to RNF4-dependent proteolytic targeting of PML. FEBS Lett 582: 3174–3178.
Xie Y, Kerscher O, Kroetz MB, McConchie HF, Sung P, Hochstrasser M . (2007). The yeast Hex3.Slx8 heterodimer is a ubiquitin ligase stimulated by substrate sumoylation. J Biol Chem 282: 34176–34184.
Yousef AF, Xu GW, Mendez M, Brandl CJ, Mymryk JS . (2008). Coactivator requirements for p53-dependent transcription in the yeast Saccharomyces cerevisiae. Int J Cancer 122: 942–946.
Zhang Z, Smith MM, Mymryk JS . (2001). Interaction of the E1A oncoprotein with Yak1p, a novel regulator of yeast pseudohyphal differentiation, and related mammalian kinases. Mol Biol Cell 12: 699–710.
Zhu S, Sachdeva M, Wu F, Lu Z, Mo YY . (2009). Ubc9 promotes breast cell invasion and metastasis in a sumoylation-independent manner. Oncogene 29: 1763–1772.
Acknowledgements
This work was supported by a grant from the Canadian Institutes of Health Research to JSM (MOP-75647). AFY and PP were supported by CIHR Strategic Training Program in Cancer Research and Technology Transfer awards. GJF and JNA held OGS and OGSST awards. We thank Drs C Brandl, R Hay, J Taylor, K Uzunova, J Dohmen, A Strunnikov, G Fink, E Yeh and O Janne for generously providing reagents essential for this study.
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Yousef, A., Fonseca, G., Pelka, P. et al. Identification of a molecular recognition feature in the E1A oncoprotein that binds the SUMO conjugase UBC9 and likely interferes with polySUMOylation. Oncogene 29, 4693–4704 (2010). https://doi.org/10.1038/onc.2010.226
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DOI: https://doi.org/10.1038/onc.2010.226
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