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

Wei Zhang; Kuen Phon Wu; Maria A. Sartori; Hari B. Kamadurai; Alban Ordureau; Chong Jiang; Peter Y. Mercredi; Ryan Murchie; Jicheng Hu; Avinash Persaud; Manjeet Mukherjee; Nan Li; Anne Doye; John R. Walker; Yi Sheng; Zhenyue Hao; Yanjun Li; Kevin R. Brown; Emmanuel Lemichez; Junjie Chen; Yufeng Tong; J. Wade Harper; Jason Moffat; Daniela Rotin; Brenda A. Schulman; Sachdev S. Sidhu

doi:10.1016/j.molcel.2016.02.005

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

Wei Zhang, Kuen Phon Wu, Maria A. Sartori, Hari B. Kamadurai, Alban Ordureau, Chong Jiang, Peter Y. Mercredi, Ryan Murchie, Jicheng Hu, Avinash Persaud, Manjeet Mukherjee, Nan Li, Anne Doye, John R. Walker, Yi Sheng, Zhenyue Hao, Yanjun Li, Kevin R. Brown, Emmanuel Lemichez, Junjie ChenYufeng Tong, J. Wade Harper, Jason Moffat, Daniela Rotin, Brenda A. Schulman, Sachdev S. Sidhu

Research output: Contribution to journal › Article › peer-review

131 Scopus citations

Abstract

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 toolkit for probing functions of HECT E3s, and establishes a general strategy for systematic development of modulators targeting families of signaling proteins.

Original language	English (US)
Pages (from-to)	121-136
Number of pages	16
Journal	Molecular cell
Volume	62
Issue number	1
DOIs	https://doi.org/10.1016/j.molcel.2016.02.005
State	Published - Apr 7 2016

ASJC Scopus subject areas

Molecular Biology
Cell Biology

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Zhang, W., Wu, K. P., Sartori, M. A., Kamadurai, H. B., Ordureau, A., Jiang, C., Mercredi, P. Y., Murchie, R., Hu, J., Persaud, A., Mukherjee, M., Li, N., Doye, A., Walker, J. R., Sheng, Y., Hao, Z., Li, Y., Brown, K. R., Lemichez, E., ... Sidhu, S. S. (2016). System-Wide Modulation of HECT E3 Ligases with Selective Ubiquitin Variant Probes. Molecular cell, 62(1), 121-136. https://doi.org/10.1016/j.molcel.2016.02.005

Zhang, W, Wu, KP, Sartori, MA, Kamadurai, HB, Ordureau, A, Jiang, C, Mercredi, PY, Murchie, R, Hu, J, Persaud, A, Mukherjee, M, Li, N, Doye, A, Walker, JR, Sheng, Y, Hao, Z, Li, Y, Brown, KR, Lemichez, E, Chen, J, Tong, Y, Harper, JW, Moffat, J, Rotin, D, Schulman, BA & Sidhu, SS 2016, 'System-Wide Modulation of HECT E3 Ligases with Selective Ubiquitin Variant Probes', Molecular cell, vol. 62, no. 1, pp. 121-136. https://doi.org/10.1016/j.molcel.2016.02.005

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abstract = "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 toolkit for probing functions of HECT E3s, and establishes a general strategy for systematic development of modulators targeting families of signaling proteins.",

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AU - Zhang, Wei

AU - Wu, Kuen Phon

AU - Sartori, Maria A.

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AU - Ordureau, Alban

AU - Jiang, Chong

AU - Mercredi, Peter Y.

AU - Murchie, Ryan

AU - Hu, Jicheng

AU - Persaud, Avinash

AU - Mukherjee, Manjeet

AU - Li, Nan

AU - Doye, Anne

AU - Walker, John R.

AU - Sheng, Yi

AU - Hao, Zhenyue

AU - Li, Yanjun

AU - Brown, Kevin R.

AU - Lemichez, Emmanuel

AU - Chen, Junjie

AU - Tong, Yufeng

AU - Harper, J. Wade

AU - Moffat, Jason

AU - Rotin, Daniela

AU - Schulman, Brenda A.

AU - Sidhu, Sachdev S.

PY - 2016/4/7

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N2 - 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 toolkit for probing functions of HECT E3s, and establishes a general strategy for systematic development of modulators targeting families of signaling proteins.

AB - 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 toolkit for probing functions of HECT E3s, and establishes a general strategy for systematic development of modulators targeting families of signaling proteins.

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

Abstract

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