title: Overexpression of a dominant-negative mutant Ubc9 is associated with increased sensitivity to anticancer drugs.
abstract: Ubc9 is an E2-conjugating enzyme required for sumoylation and has been implicated in regulating several critical cellular pathways. We have shown previously that Ubc9 is important for sumoylation and nucleolar delocalization of topoisomerase (topo) I in response to topo I inhibitors such as topotecan. However, the role for Ubc9 in tumor drug responsiveness is not clear. In this study, we found that although MCF7 cells expressing a Ubc9 dominant-negative mutant (Ubc9-DN) display decreased activity of topo I, these cells are more sensitive to the topo I inhibitor topotecan and other anticancer agents such as VM-26 and cisplatin. In addition, we found that alteration of Ubc9 expression correlates with drug responsiveness in tumor cell lines. To understand possible mechanisms of Ubc9-associated drug responsiveness, we examined several proteins that have been shown to interact with Ubc9 and that may be involved in drug responsiveness. One such protein is Daxx, which is a Fas-associated protein that plays a role in Fas-mediated apoptosis by participating in a caspase-independent pathway through activation of apoptosis signal-regulating kinase 1 and c-Jun NH(2)-terminal kinase. We found that cells expressing Ubc9-DN accumulate more cytoplasmic Daxx than the control cells. Because cytoplasmic Daxx is believed to participate in cellular apoptosis, we suggest that the interaction of Ubc9 with Daxx and subsequent alteration in the subcellular localization of Daxx may contribute to the increased sensitivity to anticancer drugs in the cells expressing Ubc9-DN. Finally, we found that overexpression of Daxx sensitizes cells to anticancer drugs possibly in part through alterations of the ratio of cytoplasmic and nuclear Daxx. Together, our results suggest a role for Ubc9 in tumor drug responsiveness.
authors.affiliation: Department of Biopharmaceutical Sciences, and the Cancer Center, University of Illinois at Chicago, Chicago, Illinois, USA. ymo@siumed.edu
authors.forename: Yin-Yuan
authors.initials: YY
authors.lastname: Mo
authors.forename: Yanni
authors.initials: Y
authors.lastname: Yu
authors.forename: P L Rachel
authors.initials: PL
authors.lastname: Ee
authors.forename: William T
authors.initials: WT
authors.lastname: Beck
mesh_terms: D048868:Adaptor Proteins, Signal Transducing; D000970:Antineoplastic Agents / Q000494:pharmacology*; D002352:Carrier Proteins / Q000096:biosynthesis / Q000378:metabolism; D045744:Cell Line, Tumor; D056970:Co-Repressor Proteins; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005799:Genes, Dominant; D006367:HeLa Cells; D006801:Humans; D047908:Intracellular Signaling Peptides and Proteins; D018832:Molecular Chaperones; D009687:Nuclear Proteins / Q000096:biosynthesis / Q000378:metabolism; D013347:Subcellular Fractions / Q000378:metabolism; D013713:Teniposide / Q000494:pharmacology; D059004:Topoisomerase I Inhibitors; D059005:Topoisomerase II Inhibitors; D019772:Topotecan / Q000494:pharmacology; D044763:Ubiquitin-Conjugating Enzymes / Q000096:biosynthesis / Q000235:genetics / Q000502:physiology*; D000098843:Ubiquitin-Conjugating Enzyme UBC9
chemical_list: D048868:Adaptor Proteins, Signal Transducing; D000970:Antineoplastic Agents; D002352:Carrier Proteins; D056970:Co-Repressor Proteins; C106985:DAXX protein, human; D004791:Enzyme Inhibitors; D047908:Intracellular Signaling Peptides and Proteins; D018832:Molecular Chaperones; D009687:Nuclear Proteins; D059004:Topoisomerase I Inhibitors; D059005:Topoisomerase II Inhibitors; D019772:Topotecan; D013713:Teniposide; D044763:Ubiquitin-Conjugating Enzymes; D000098843:Ubiquitin-Conjugating Enzyme UBC9