The regulatory process known as protein ubiquitination modifies cellular proteins with far-reaching impacts on human health and disease. It is involved in every known biological process and is implicated in a growing range of diseases that includes cancers, neurodegenerative diseases, muscle wasting, etc. We are studying the expanding roles of protein ubiquitination to understand underlying principles of the process that can guide future efforts to manipulate or target the process. Over its thirteen-year course, the scope of the project has expanded beyond the single ubiquitin E3 ligase, the breast cancer susceptibility protein, BRCA1/BARD1, and its E2 conjugating enzymes. To reflect our expanding field of investigation and the expanding functionalities of E2s and E3s we have uncovered, the project is renamed """"""""Expanding Roles of E2 and E3 enzymes in Ubiquitin Transfer."""""""" In Aim 1, we will use new mechanistic insights and new technology to generate RING E3s with enhanced activities for use in functional studies.
In Aim 2, we will investigate the RING-Between-RING E3s, newly identified as a novel RING-HECT hybrid class of E3s.
In Aim 3, we seek to define novel mechanisms used by E2s.

Public Health Relevance

The regulatory process known as protein ubiquitination modifies cellular proteins with far---reaching impacts on human health and disease. It is involved in every known biological process and is implicated in a growing range of diseases that includes cancers, neurodegenerative diseases, muscle wasting, etc. We are studying the expanding roles of protein ubiquitination to understand underlying principles of the process that can guide future efforts to manipulate or target the process.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM088055-15
Application #
8710250
Study Section
Membrane Biology and Protein Processing (MBPP)
Program Officer
Wehrle, Janna P
Project Start
1998-12-21
Project End
2017-04-30
Budget Start
2014-05-01
Budget End
2015-04-30
Support Year
15
Fiscal Year
2014
Total Cost
$528,224
Indirect Cost
$178,224
Name
University of Washington
Department
Biochemistry
Type
Schools of Medicine
DUNS #
605799469
City
Seattle
State
WA
Country
United States
Zip Code
98195
Dove, Katja K; Stieglitz, Benjamin; Duncan, Emily D et al. (2016) Molecular insights into RBR E3 ligase ubiquitin transfer mechanisms. EMBO Rep 17:1221-35
Dove, Katja K; Klevit, Rachel E; Rittinger, Katrin (2015) pUBLically unzipping Parkin: how phosphorylation exposes a ligase bit by bit. EMBO J 34:2486-8
Wu, Wenwen; Nishikawa, Hiroyuki; Fukuda, Takayo et al. (2015) Interaction of BARD1 and HP1 Is Required for BRCA1 Retention at Sites of DNA Damage. Cancer Res 75:1311-21
Vittal, Vinayak; Stewart, Mikaela D; Brzovic, Peter S et al. (2015) Regulating the Regulators: Recent Revelations in the Control of E3 Ubiquitin Ligases. J Biol Chem 290:21244-51
Vittal, Vinayak; Shi, Lei; Wenzel, Dawn M et al. (2015) Intrinsic disorder drives N-terminal ubiquitination by Ube2w. Nat Chem Biol 11:83-9
Metzger, Meredith B; Pruneda, Jonathan N; Klevit, Rachel E et al. (2014) RING-type E3 ligases: master manipulators of E2 ubiquitin-conjugating enzymes and ubiquitination. Biochim Biophys Acta 1843:47-60
Pruneda, Jonathan N; Smith, F Donelson; Daurie, Angela et al. (2014) E2~Ub conjugates regulate the kinase activity of Shigella effector OspG during pathogenesis. EMBO J 33:437-49
Warfield, Linda; Tuttle, Lisa M; Pacheco, Derek et al. (2014) A sequence-specific transcription activator motif and powerful synthetic variants that bind Mediator using a fuzzy protein interface. Proc Natl Acad Sci U S A 111:E3506-13
Soss, Sarah E; Klevit, Rachel E; Chazin, Walter J (2013) Activation of UbcH5c~Ub is the result of a shift in interdomain motions of the conjugate bound to U-box E3 ligase E4B. Biochemistry 52:2991-9
Starita, Lea M; Pruneda, Jonathan N; Lo, Russell S et al. (2013) Activity-enhancing mutations in an E3 ubiquitin ligase identified by high-throughput mutagenesis. Proc Natl Acad Sci U S A 110:E1263-72

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