Structural insights into ADP-ribosylation of ubiquitin by Deltex family E3 ubiquitin ligases

TY - JOUR

T1 - Structural insights into ADP-ribosylation of ubiquitin by Deltex family E3 ubiquitin ligases

AU - Chatrin, Chatrin

AU - Gabrielsen, Mads

AU - Buetow, Lori

AU - Nakasone, Mark A.

AU - Ahmed, Syed F.

AU - Sumpton, David

AU - Sibbet, Gary J.

AU - Smith, Brian O.

AU - Huang, Danny T.

N1 - Copyright © 2020 The Authors, some rights reserved; exclusive licensee American Association for the Advancement of Science. No claim to original U.S. Government Works. Distributed under a Creative Commons Attribution NonCommercial License 4.0 (CC BY-NC).

PY - 2020/9

Y1 - 2020/9

N2 - Cellular cross-talk between ubiquitination and other posttranslational modifications contributes to the regulation of numerous processes. One example is ADP-ribosylation of the carboxyl terminus of ubiquitin by the E3 DTX3L/ADP-ribosyltransferase PARP9 heterodimer, but the mechanism remains elusive. Here, we show that independently of PARP9, the conserved carboxyl-terminal RING and DTC (Deltex carboxyl-terminal) domains of DTX3L and other human Deltex proteins (DTX1 to DTX4) catalyze ADP-ribosylation of ubiquitin's Gly76. Structural studies reveal a hitherto unknown function of the DTC domain in binding NAD+ Deltex RING domain recruits E2 thioesterified with ubiquitin and juxtaposes it with NAD+ bound to the DTC domain to facilitate ADP-ribosylation of ubiquitin. This ubiquitin modification prevents its activation but is reversed by the linkage nonspecific deubiquitinases. Our study provides mechanistic insights into ADP-ribosylation of ubiquitin by Deltex E3s and will enable future studies directed at understanding the increasingly complex network of ubiquitin cross-talk.

AB - Cellular cross-talk between ubiquitination and other posttranslational modifications contributes to the regulation of numerous processes. One example is ADP-ribosylation of the carboxyl terminus of ubiquitin by the E3 DTX3L/ADP-ribosyltransferase PARP9 heterodimer, but the mechanism remains elusive. Here, we show that independently of PARP9, the conserved carboxyl-terminal RING and DTC (Deltex carboxyl-terminal) domains of DTX3L and other human Deltex proteins (DTX1 to DTX4) catalyze ADP-ribosylation of ubiquitin's Gly76. Structural studies reveal a hitherto unknown function of the DTC domain in binding NAD+ Deltex RING domain recruits E2 thioesterified with ubiquitin and juxtaposes it with NAD+ bound to the DTC domain to facilitate ADP-ribosylation of ubiquitin. This ubiquitin modification prevents its activation but is reversed by the linkage nonspecific deubiquitinases. Our study provides mechanistic insights into ADP-ribosylation of ubiquitin by Deltex E3s and will enable future studies directed at understanding the increasingly complex network of ubiquitin cross-talk.

KW - ADP-Ribosylation

KW - Humans

KW - NAD/metabolism

KW - Ubiquitin/metabolism

KW - Ubiquitin-Protein Ligases/metabolism

KW - Ubiquitination

U2 - 10.1126/sciadv.abc0418

DO - 10.1126/sciadv.abc0418

M3 - Article

C2 - 32948590

SN - 2375-2548

VL - 6

JO - Science Advances

JF - Science Advances

IS - 38

M1 - eabc0418

ER -