10779/crick.12707966.v1
A Barbara Dirac-Svejstrup
A Barbara
Dirac-Svejstrup
Jane Walker
Jane
Walker
Peter Faull
Peter
Faull
Vesela Encheva
Vesela
Encheva
Vyacheslav Akimov
Vyacheslav
Akimov
Michele Puglia
Michele
Puglia
David Perkins
David
Perkins
Sandra Kümper
Sandra
Kümper
Suchete S Hunjan
Suchete S
Hunjan
Blagoy Blagoev
Blagoy
Blagoev
Ambrosius P Snijders
Ambrosius P
Snijders
David J Powell
David J
Powell
Jesper Q Svejstrup
Jesper Q
Svejstrup
DDI2 Is a ubiquitin-directed endoprotease responsible for cleavage of transcription factor NRF1.
The Francis Crick Institute
2020
Bortezomib
DDI2
Ddi1
NFE2L1
NRF1
proteasome
proteasome inhibition
ubiquitin
ubiquitin protease
Svejstrup FC001166
PRT
CS-ack
FC-ack
HTS-ack
GSK
Developmental Biology
06 Biological Sciences
11 Medical and Health Sciences
2020-07-24 13:23:48
Journal contribution
https://crick.figshare.com/articles/journal_contribution/DDI2_Is_a_ubiquitin-directed_endoprotease_responsible_for_cleavage_of_transcription_factor_NRF1_/12707966
The Ddi1/DDI2 proteins are ubiquitin shuttling factors, implicated in a variety of cellular functions. In addition to ubiquitin-binding and ubiquitin-like domains, they contain a conserved region with similarity to retroviral proteases, but whether and how DDI2 functions as a protease has remained unknown. Here, we show that DDI2 knockout cells are sensitive to proteasome inhibition and accumulate high-molecular weight, ubiquitylated proteins that are poorly degraded by the proteasome. These proteins are targets for the protease activity of purified DDI2. No evidence for DDI2 acting as a de-ubiquitylating enzyme was uncovered, which could suggest that it cleaves the ubiquitylated protein itself. In support of this idea, cleavage of transcription factor NRF1 is known to require DDI2 activity in vivo. We show that DDI2 is indeed capable of cleaving NRF1 in vitro but only when NRF1 protein is highly poly-ubiquitylated. Together, these data suggest that DDI2 is a ubiquitin-directed endoprotease.