Abstract
Rad23 is a highly conserved protein involved in nucleotide excision repair (NER) that associates with the proteasome via its N-terminus. Its C-terminal ubiquitin-associated (UBA) domain is evolutionarily conserved from yeast to humans. However, the cellular function of UBA domains is not completely understood. Recently, RAD23 and DDI1, both DNA damage-inducible genes encoding proteins with UBA domains, were implicated genetically in Pds1-dependent mitotic control in yeast. The UBA domains of RAD23 and DDI1 are required for these interactions. Timely degradation of Pds1 via the ubiquitin/proteasome pathway allows anaphase onset and is crucial for chromosome maintenance. Here, we show that Rad23 and Ddi1 interact directly with ubiquitin and that this interaction is dependent on their UBA domains, providing a possible mechanism for UBA-dependent cell cycle control. Moreover, we show that a hydrophobic surface on the UBA domain, which from structural work had been predicted to be a protein–protein interaction interface, is indeed required for ubiquitin binding. By demonstrating that UBA domains interact with ubiquitin, we have provided the first indication of a cellular function for the UBA domain.
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References
Hofmann, K. & Bucher, P. Trends Biochem. Sci. 21, 172–173 (1996).
Dieckmann, T. et al. Nature Struct. Biol. 5, 1042–1047 (1998).
Masutani, C. et al. Mol. Cell. Biol. 17, 6915–6923 (1997).
Sugasawa, K. et al. Mol. Cell. Biol. 17, 6924–6931 (1997).
Schauber, C. et al. Nature 391, 715–718 (1998).
Watkins, J.F., Sung, P., Prakash, L. & Prakash, S. Mol. Cell. Biol. 13, 7757–7765 (1993).
Hiyama, H. et al. J. Biol. Chem. 274, 28019–28025 (1999).
Miao, F. et al. J. Biol. Chem. 275, 28433–28438 (2000).
Withers-Ward, E.S. et al. J. Virol. 71, 9732–9742 (1997).
Liu, N. et al. Proc. Natl. Acad. Sci. USA 94, 9232–9237 (1997).
Liu, Y., Dai, H. & Xiao, W. Mol. Gen. Genet. 255, 533–542 (1997).
Lustgarten, V. & Gerst, J.E. Mol. Cell Biol. 19, 4480–4494 (1999).
Uetz, P. et al. Nature 403, 623–627 (2000).
Yamamoto, A., Guacci, V. & Koshland, D. J. Cell. Biol. 133, 99–110 (1996).
Yamamoto, A., Guacci, V. & Koshland, D. J. Cell. Biol. 133, 85–97 (1996).
Cohen-Fix, O., Peters, J.M., Kirschner, M.W. & Koshland, D. Genes Dev. 10, 3081–3093 (1996).
Clarke, D.J. et al. Mol. Cell Biol. 21, 1997–2007 (2001).
Lindsey, D.F. et al. J. Biol. Chem. 273, 29178–29187 (1998).
Vadlamudi, R.K., Joung, I., Strominger, J.L. & Shin, J. J. Biol. Chem. 271, 20235–20237 (1996).
Schultz, J., Milpetz, F., Bork, P. & Ponting, C.P. Proc. Natl. Acad. Sci. USA 95, 5857–5864 (1998).
Joazeiro, C.A. et al. Science 286, 309–312 (1999).
Ortolan, T.G. et al. Nature Cell Biol. 2, 601–608 (2000).
Richardson, H.E., Wittenberg, C., Cross, F.R. & Reed, S.I. Cell 59, 1127–1133 (1989).
Clarke, D.J., Segal, M., Mondesert, G. & Reed, S.I. Curr. Biol. 9, 365–368 (1999).
Sherman, F., Fink, G. & Hicks, J.B. Methods in yeast genetics (Cold Spring Harbor Laboratory, Cold Spring Harbor, New York; 1982).
Wach, A., Brachat, A., Pohlmann, R. & Philippsen, P. Yeast 10, 1793–1808 (1994).
Mondesert, G., Clarke, D.J. & Reed, S.I. Genetics 147, 421–434 (1997).
Gyuris, J., Golemis, E., Chertkov, H. & Brent, R. Cell 75, 791–803 (1993).
Simon, M.N. et al. Nature 376, 702–705 (1995).
Muhlrad, D., Hunter, R. & Parker, R. Yeast 8, 79–82 (1992).
Kaiser, P., Sia, R.A., Bardes, E.G., Lew, D.J. & Reed, S.I. Genes Dev. 12, 2587–2597 (1998).
Heitz, F. et al. Biochemistry 36, 4995–5003 (1997).
Acknowledgements
B.L.B. was supported by both NIH and University of California Office of the President fellowships. D.J.C. was supported by both EMBO and DOD Breast Cancer Research Program fellowships. We would like to thank L. Prakash for the α-Rad23 antibody and E. Komives for help in rendering the ribbon diagram of the HHR23A UBA2 NMR structure. We would also like to thank the members of the Reed laboratory and the Scripps Cell Cycle groups for helpful discussions and specifically J. Tainer for use of his laboratory and computing facilities.
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Bertolaet, B., Clarke, D., Wolff, M. et al. UBA domains of DNA damage-inducible proteins interact with ubiquitin. Nat Struct Mol Biol 8, 417–422 (2001). https://doi.org/10.1038/87575
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DOI: https://doi.org/10.1038/87575
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