System-Wide Modulation of HECT E3 Ligases with Selective Ubiquitin Variant Probes

被引:130
作者
Zhang, Wei [1 ]
Wu, Kuen-Phon [2 ]
Sartori, Maria A. [1 ]
Kamadurai, Hari B. [2 ,14 ]
Ordureau, Alban [3 ]
Jiang, Chong [4 ,5 ]
Mercredi, Peter Y. [2 ]
Murchie, Ryan [4 ,5 ]
Hu, Jicheng [6 ]
Persaud, Avinash [4 ,5 ]
Mukherjee, Manjeet [2 ,15 ]
Li, Nan [7 ]
Doye, Anne [8 ]
Walker, John R. [6 ]
Sheng, Yi [9 ]
Hao, Zhenyue [10 ]
Li, Yanjun [6 ]
Brown, Kevin R. [1 ]
Lemichez, Emmanuel [8 ]
Chen, Junjie [7 ]
Tong, Yufeng [6 ,11 ]
Harper, J. Wade [3 ]
Moffat, Jason [1 ,12 ]
Rotin, Daniela [4 ,5 ]
Schulman, Brenda A. [2 ,13 ]
Sidhu, Sachdev S. [1 ,12 ]
机构
[1] Univ Toronto, Banting & Best Dept Med Res, Donnelly Ctr Cellular & Biomol Res, 160 Coll St, Toronto, ON M5S3E1, Canada
[2] St Jude Childrens Res Hosp, Dept Biol Struct, 332 N Lauderdale St, Memphis, TN 38105 USA
[3] Harvard Univ, Sch Med, Dept Cell Biol, Boston, MA 02115 USA
[4] Univ Toronto, Hosp Sick Children, Cell Biol Program, Toronto, ON M5G 0A4, Canada
[5] Univ Toronto, Dept Biochem, Toronto, ON M5G 0A4, Canada
[6] Univ Toronto, Struct Genom Consortium, Toronto, ON M5G1LP, Canada
[7] Univ Texas MD Anderson Canc Ctr, Dept Expt Radiat Oncol, 1515 Holcombe Blvd, Houston, TX 77030 USA
[8] Univ Nice Sophia Antipolis, Equipe Labellisee Ligue Contre Canc, Ctr Mediterraneen Med Mol, INSERM,U1065,C3M, 151 Route St Antoine Ginestiere,BP 2 3194, F-23194 Nice, France
[9] York Univ, Dept Biol, 4700 Keele St, Toronto, ON M3J 1P3, Canada
[10] Univ Hlth Network, Campbell Family Canc Res Inst, Toronto, ON M5G2C1, Canada
[11] Univ Toronto, Dept Pharmacol & Toxicol, Toronto, ON M5G 1L7, Canada
[12] Univ Toronto, Kings Coll Cir 1, Dept Mol Genet, 100 Coll St, Toronto, ON M5S 1A8, Canada
[13] St Jude Childrens Res Hosp, Howard Hughes Med Inst, 332 N Lauderdale St, Memphis, TN 38105 USA
[14] Biogen Inc, 161 First St, Cambridge, MA 02142 USA
[15] Univ Leicester, Dept Mol & Cell Biol, Leicester LE1 9HN, Leics, England
基金
巴西圣保罗研究基金会; 加拿大创新基金会; 英国惠康基金;
关键词
BREAST-CANCER CELLS; SMALL-MOLECULE; TARGETING CANCER; BINDING; INHIBITORS; MIGRATION; MECHANISM; PROTEINS; COMPLEX; FAMILY;
D O I
10.1016/j.molcel.2016.02.005
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
HECT-family E3 ligases ubiquitinate protein substrates to control virtually every eukaryotic process and are misregulated in numerous diseases. Nonetheless, understanding of HECT E3s is limited by a paucity of selective and potent modulators. To overcome this challenge, we systematically developed ubiquitin variants (UbVs) that inhibit or activate HECT E3s. Structural analysis of 6 HECT-UbV complexes revealed UbV inhibitors hijacking the E2-binding site and activators occupying a ubiquitin-binding exosite. Furthermore, UbVs unearthed distinct regulation mechanisms among NEDD4 subfamily HECTs and proved useful for modulating therapeutically relevant targets of HECT E3s in cells and intestinal organoids, and in a genetic screen that identified a role for NEDD4L in regulating cell migration. Our work demonstrates versatility of UbVs for modulating activity across an E3 family, defines mechanisms and provides a tool-kit for probing functions of HECT E3s, and establishes a general strategy for systematic development of modulators targeting families of signaling proteins.
引用
收藏
页码:121 / 136
页数:16
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