1yrv Citations

A human ubiquitin conjugating enzyme (E2)-HECT E3 ligase structure-function screen.


Here we describe a systematic structure-function analysis of the human ubiquitin (Ub) E2 conjugating proteins, consisting of the determination of 15 new high-resolution three-dimensional structures of E2 catalytic domains, and autoubiquitylation assays for 26 Ub-loading E2s screened against a panel of nine different HECT (homologous to E6-AP carboxyl terminus) E3 ligase domains. Integration of our structural and biochemical data revealed several E2 surface properties associated with Ub chain building activity; (1) net positive or neutral E2 charge, (2) an "acidic trough" located near the catalytic Cys, surrounded by an extensive basic region, and (3) similarity to the previously described HECT binding signature in UBE2L3 (UbcH7). Mass spectrometry was used to characterize the autoubiquitylation products of a number of functional E2-HECT pairs, and demonstrated that HECT domains from different subfamilies catalyze the formation of very different types of Ub chains, largely independent of the E2 in the reaction. Our data set represents the first comprehensive analysis of E2-HECT E3 interactions, and thus provides a framework for better understanding the molecular mechanisms of ubiquitylation.

Articles - 1yrv mentioned but not cited (1)

  1. A human ubiquitin conjugating enzyme (E2)-HECT E3 ligase structure-function screen. Sheng Y, Hong JH, Doherty R, Srikumar T, Shloush J, Avvakumov GV, Walker JR, Xue S, Neculai D, Wan JW, Kim SK, Arrowsmith CH, Raught B, Dhe-Paganon S. Mol. Cell Proteomics 11 329-341 (2012)

Reviews citing this publication (9)

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Articles citing this publication (49)

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  14. Molecular Characterization of LubX: Functional Divergence of the U-Box Fold by Legionella pneumophila. Quaile AT, Urbanus ML, Stogios PJ, Nocek B, Skarina T, Ensminger AW, Savchenko A. Structure 23 1459-1469 (2015)
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  19. Structure of UBE2Z Enzyme Provides Functional Insight into Specificity in the FAT10 Protein Conjugation Machinery. Schelpe J, Monté D, Dewitte F, Sixma TK, Rucktooa P. J. Biol. Chem. 291 630-639 (2016)
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  21. Crystal Structure of a Ube2S-Ubiquitin Conjugate. Lorenz S, Bhattacharyya M, Feiler C, Rape M, Kuriyan J. PLoS ONE 11 e0147550 (2016)
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  32. An E2-guided E3 Screen Identifies the RNF17-UBE2U Pair as Regulator of the Radiosensitivity, Immunodeficiency, Dysmorphic Features, and Learning Difficulties (RIDDLE) Syndrome Protein RNF168. Guo Y, An L, Ng HM, Sy SM, Huen MS. J. Biol. Chem. 292 967-978 (2017)
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  44. Hepatitis C Virus Downregulates Ubiquitin-Conjugating Enzyme E2S Expression To Prevent Proteasomal Degradation of NS5A, Leading to Host Cells More Sensitive to DNA Damage. Pham HT, Nguyen TTT, Nguyen LP, Han SS, Lim YS, Hwang SB. J. Virol. 93 (2019)
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  49. The S. Typhi effector StoD is an E3/E4 ubiquitin ligase which binds K48- and K63-linked diubiquitin. McDowell MA, Byrne AM, Mylona E, Johnson R, Sagfors A, Crepin VF, Lea S, Frankel G. Life Sci Alliance 2 (2019)