Two distinct types of E3 ligases work in unison to regulate substrate ubiquitylation

Daniel C. Scott, David Y. Rhee, David M. Duda, Ian R. Kelsall, Jennifer L. Olszewski, Joao A. Paulo, Annemieke de Jong, Huib Ovaa, Arno F. Alpi, J. Wade Harper (Lead / Corresponding author), Brenda A. Schulman (Lead / Corresponding author)

Research output: Contribution to journalArticlepeer-review

157 Citations (Scopus)


Hundreds of human cullin-RING E3 ligases (CRLs) modify thousands of proteins with ubiquitin (UB) to achieve vast regulation. Current dogma posits that CRLs first catalyze UB transfer from an E2 to their client substrates and subsequent polyubiquitylation from various linkage-specific E2s. We report an alternative E3-E3 tagging cascade: many cellular NEDD8-modified CRLs associate with a mechanistically distinct thioester-forming RBR-type E3, ARIH1, and rely on ARIH1 to directly add the first UB and, in some cases, multiple additional individual monoubiquitin modifications onto CRL client substrates. Our data define ARIH1 as a component of the human CRL system, demonstrate that ARIH1 can efficiently and specifically mediate monoubiquitylation of several CRL substrates, and establish principles for how two distinctive E3s can reciprocally control each other for simultaneous and joint regulation of substrate ubiquitylation. These studies have broad implications for CRL-dependent proteostasis and mechanisms of E3-mediated UB ligation.

Original languageEnglish
Pages (from-to)1198-1214.e24
Number of pages41
Issue number5
Publication statusPublished - 25 Aug 2016


  • Journal article


Dive into the research topics of 'Two distinct types of E3 ligases work in unison to regulate substrate ubiquitylation'. Together they form a unique fingerprint.

Cite this