Abstract

Theregulatoryprocessknownasproteinubiquitinationmodifiescellularproteinswith far-reachingimpactsonhumanhealthanddisease.Itisinvolvedineveryknown biologicalprocessandisimplicatedinagrowingrangeofdiseasesthatincludes cancers,neurodegenerativediseases,musclewasting,etc.Studiesconducted worldwideoverthepasttwentyyearshavedefinedthemechanismsbywhichprotein ubiquitinationiscarriedout,withoneobviousexception.Almostlittleisknownabouthow substrates(i.e.,thecellularproteinsthataremodifiedbythissystem)arerecognized andhowtheyfitintotheultimatemechanism.Inthecominggrantperiod,weproposeto rectifythissituationfortwoimportantubiquitinE3ligases,BRCA1/BARD1andHHARI. Ourgoalsaremadepossiblebyseveralimportantadvances,includingdefinitionofbona fidesubstratesforeachoftheseE3sandtheabilitytochemicallysynthesize biochemicallyhomogeneousversionoftheproducts.Wewillalsoinvestigatetwo uncharacterizedhumanE2Ubiquitin-conjugatingenzymesthateachhave unusual/uniqueactivitiesinthattheyattachUbtoproteingroupsotherthanlysines, therebyexpandingtheuniverseofpossibleattachmentsites.Wewilluseacombination ofbiochemical,structural,andcellularapproachestoaddressquestionsoffundamental importanceandgeneralrelevance.

Public Health Relevance

Theregulatoryprocessknownasproteinubiquitinationmodifiescellularproteins withfar-??reachingimpactsonhumanhealthanddisease.Itisinvolvedinevery knownbiologicalprocessandisimplicatedinagrowingrangeofdiseasesthat includescancers,neurodegenerativediseases,musclewasting,etc.Wearestudying theexpandingrolesofproteinubiquitinationtounderstandunderlyingprinciplesof theprocessthatcanguidefutureeffortstomanipulateortargettheprocess.

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
3R01GM088055-19S1
Application #
9881170
Study Section
Program Officer
Cubano, Luis Angel
Project Start
1998-12-21
Project End
2021-08-31
Budget Start
2018-09-01
Budget End
2019-08-31
Support Year
19
Fiscal Year
2019
Total Cost
Indirect Cost

  Institution

Name
University of Washington
Department
Biochemistry
Type
Schools of Medicine
DUNS #
605799469
City
Seattle
State
WA
Country
United States
Zip Code
98195

  Publications

Akturk, Anil; Wasilko, David J; Wu, Xiaochun et al. (2018) Mechanism of phosphoribosyl-ubiquitination mediated by a single Legionella effector. Nature 557:729-733
DaRosa, Paul A; Harrison, Joseph S; Zelter, Alex et al. (2018) A Bifunctional Role for the UHRF1 UBL Domain in the Control of Hemi-methylated DNA-Dependent Histone Ubiquitylation. Mol Cell 72:753-765.e6
Stewart, Mikaela D; Zelin, Elena; Dhall, Abhinav et al. (2018) BARD1 is necessary for ubiquitylation of nucleosomal histone H2A and for transcriptional regulation of estrogen metabolism genes. Proc Natl Acad Sci U S A 115:1316-1321
Pierce, Sarah B; Stewart, Mikaela D; Gulsuner, Suleyman et al. (2018) De novo mutation in RING1 with epigenetic effects on neurodevelopment. Proc Natl Acad Sci U S A 115:1558-1563
Dove, Katja K; Olszewski, Jennifer L; Martino, Luigi et al. (2017) Structural Studies of HHARI/UbcH7?Ub Reveal Unique E2?Ub Conformational Restriction by RBR RING1. Structure 25:890-900.e5
Stewart, Mikaela D; Duncan, Emily D; Coronado, Ernesto et al. (2017) Tuning BRCA1 and BARD1 activity to investigate RING ubiquitin ligase mechanisms. Protein Sci 26:475-483
Dove, Katja K; Kemp, Hilary A; Di Bona, Kristin R et al. (2017) Two functionally distinct E2/E3 pairs coordinate sequential ubiquitination of a common substrate in Caenorhabditis elegans development. Proc Natl Acad Sci U S A 114:E6576-E6584
Dove, Katja K; Klevit, Rachel E (2017) RING-Between-RING E3 Ligases: Emerging Themes amid the Variations. J Mol Biol 429:3363-3375
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
Harrison, Joseph S; Cornett, Evan M; Goldfarb, Dennis et al. (2016) Hemi-methylated DNA regulates DNA methylation inheritance through allosteric activation of H3 ubiquitylation by UHRF1. Elife 5:

Showing the most recent 10 out of 31 publications