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

来源：

MOLECULAR CELL | 2016年 / 62卷 / 01期

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

页码：121 / 136

页数：16

共 59 条

[11] Activation of the E3 ubiquitin ligase Itch through a phosphorylation-induced conformational change [J].