User:Jeffrey D. Singer
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- Jeffrey D. Singer, Ph.D.
- Portland State University
- Department of Biology
- P.O. Box 751
- Portland, OR 97207
- Office: U62A SB2
- Lab: L31 SB2
Laboratory for Ubiquitin Signaling
- 503-725-8742 (office)
- 503-725-2496 (lab)
- 503-725-3888 (fax)
- 503-896-6704 (cell)
- 1995, PhD, University of Utah
- 1987, MS, University of New Mexico
- 1983, BS, University of Oregon
In order to gain a better understanding of how ubiquitin-signaling is regulated I have focused on the structure and function of an E3 ligase that I identified as a regulator of a cyclin called cyclin E. This cyclin is an important mediator of entrance into the cell cycle and thus is important in normal proliferative processes such as wound healing, liver regeneration as well as abnormal processes such as cancer. We have taken a broad approach that encompasses biochemical methods, proteomics, as well as mouse disease models to determine how E3 ligases work. In doing so we have uncovered unique in vivo roles for this E3 ligase as well as new molecular details regarding the structure of the active complex. These details we have uncovered regarding the structure and function of E3 ligases have allowed us to begin to assemble all of the components of the ligase to develop an in vitro system. Such a system will be useful to determine precise biochemical details about substrate selection and ubiquitin transfer to substrates. The mouse model we have developed also allows us to test hypotheses developed using biochemistry in the in vivo context.
- 1. Bear, D.G., Andrews, C.L., Singer, J.D., Morgan, W.D., Grant, R.A., von Hippel, P.H., and Platt, T. (1985). Escherichia coli transcription factor Rho has a two-domain structure in its activated form. Proc. Nat'l. Acad. Sci. USA 82: 1911-1915.
- 2. Nickoloff, J.A., Singer, J.D., Hoekstra, M.F., and Heffron, F. (1989). Double-strand Breaks Stimulate Alternative Mechanisms of Recombinational Repair. J.Mol.Biol. 207: 527-541.
- 3. Nickoloff, J.A., Singer, J.D., and Heffron, F. (1990). In vivo Analysis of the Yeast HO Nuclease Recognition Site By Site-Directed Mutagenesis. Mol. Cell Biol. 10(3): 1174-1179.
- 4. Hoekstra, M.F., Burbee, D.G., Singer, J.D., Mull, E., Chiao, E., and Heffron, F. (1991). A Tn3 derivative that can be used to make short in-frame insertions within genes. Proc. Nat'l. Acad Sci. USA 88: 5457-5461.
- 5. Bass, B.L., Hurst, S.R., and Singer, J.D. (1994). Binding properties of newly identified Xenopus proteins containing dsRNA-binding motifs. Current Biology 4(4): 301-314.
- 6. Singer, J.D., Manning, B.E., and Formosa, T. (1996). Coordinating DNA replication to produce one copy of the genome requires genes that act in ubiquitin metabolism. Mol. Cell Biol. 16(4): 1356-1366.
- 7. Singer, J.D., Gurian-West, M., Clurman, B., and Roberts, J.M. (1999). Cullin-3 targets cyclin E for ubiquitination and controls S phase in mammalian cells. Genes Dev. 13(18): 2375-87. (Featured on cover).
- 8. Sheaff, R.J., Singer, J.D., Swanger, J., Smitherman, M., Roberts, J.M., and Clurman, B.E. (2000). Proteasomal Turnover of p21Cip1 Does Not Require p21Cip1 Ubiquitination. Mol. Cell 5(2): 403-410.
- 9. Welker, M., Singer, J.D., Loeb, K., Grim, J., Bloecher, A., Gurian-West, M., Clurman, B., and Roberts, J.M. (2003). Multisite Phosphorylation by Cdk2 and GSK-3 Control Cyclin E Degradation. Mol. Cell. 12: 381-392
- 10. Klysik, J. and Singer, J.D. (2005). Mice with the Enhanced Green Flourescent Protein Gene Knocked-in to Chromosome 11 Exhibit Normal Transmission Ratios. Biochemical Genetics. 43(5/6): 321-333.
- 11. Salinas, G., Blair, L.A.C., Needleman, L.A., Gonzales, J.D., Chen, Y., Li, M., Singer, J.D. and Marshall, J. (2006) Actinfilin is a Cul3 substrate adaptor, linking GluR6 Kainate receptor subunits to the ubiquitin-proteasome pathway. J. Biol. Chem. 281(52): 40164-40173.
- 12. Wimuttisuk, W. and Singer, J.D. (2007). The Cullin3 Ubiquitin Ligase Functions As A Nedd8-Bound Heterodimer. Mol. Biol. Cell. 18: 899-909.
- 13. McEvoy, J.D., Kossatz, U., Malek, N., and Singer, J.D. (2007). Constitutive turnover of cyclin E by Cul3 maintains quiescence. Mol. Cell Biol. 27(10): 3651-3666.
- 14. Watai, Y., Kobayashi, A., Nagase, H., Mizukami, M., McEvoy, J. D., Singer, J. D., Itoh, K. and Yamamoto, M. (2007). Subcellular Localization and Cytoplasmic Complex Status of Endogenous Keap1. Genes to Cells, 12(10): 1163-1178. (Featured on cover).
- 15. Marshall, J., Blair, L. A. C., and Singer, J.D. (2009) BTB-Kelch Proteins and Ubiquitination of Kainate Receptors. in Kainate Receptors (Rodriquez-Moreno, A., and Sihra, T. eds.), Landes Scientific (in press).
- 16. Cummings, C., Bentley, C.A., Perdue, S.A., Baas, P., and Singer, J.D. (2009). The Cul3/KLHDC5 E3 Ligase Regulates P60/Katanin And Is Required For Normal Mitosis In Mammalian Cells. J. Biol. Chem., 284(17): 11663-75.
- 17. Plafker, K.S., Singer, J.D., and Plafker, S.M. (2009) The ubiquitin conjugating enzyme, UbcM2 , engages in novel interactions with components of cullin-3 based E3 ligases. Biochem., 48(15): 3527-37.