Oxygen-dependent asparagine hydroxylation of the ubiquitin-associated (UBA) domain in Cezanne regulates ubiquitin binding

Deubiquitinases (DUBs) are vital for the regulation of ubiquitin signals, and both catalytic activity of and target recruitment by DUBs need to be tightly controlled. Here, we identify asparagine hydroxylation as a novel posttranslational modification involved in the regulation of Cezanne (also know...

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Bibliographic Details
Published in:The Journal of biological chemistry 2020-02, Vol.295 (8), p.2160-2174
Main Authors: Mader, Julia, Huber, Jessica, Bonn, Florian, Dötsch, Volker, Rogov, Vladimir V., Bremm, Anja
Format: Article
Language:English
Subjects:
Amino Acid Sequence
Asparagine - metabolism
Cezanne
Consensus Sequence
deubiquitinase (DUB)
deubiquitylation (deubiquitination)
Editors' Picks
Endopeptidases - chemistry
Endopeptidases - metabolism
FIH1
HEK293 Cells
Humans
Hydroxylation
Mixed Function Oxygenases - metabolism
nuclear magnetic resonance (NMR)
OTU domain-containing protein 7B (OTUD7B)
Oxygen - metabolism
Polyubiquitin - metabolism
posttranslational modification (PTM)
Protein Binding
Protein Domains
protein-protein interaction
Repressor Proteins - metabolism
Structure-Activity Relationship
UBA domain
ubiquitin
Ubiquitin - metabolism
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Summary:Deubiquitinases (DUBs) are vital for the regulation of ubiquitin signals, and both catalytic activity of and target recruitment by DUBs need to be tightly controlled. Here, we identify asparagine hydroxylation as a novel posttranslational modification involved in the regulation of Cezanne (also known as OTU domain–containing protein 7B (OTUD7B)), a DUB that controls key cellular functions and signaling pathways. We demonstrate that Cezanne is a substrate for factor inhibiting HIF1 (FIH1)- and oxygen-dependent asparagine hydroxylation. We found that FIH1 modifies Asn35 within the uncharacterized N-terminal ubiquitin-associated (UBA)-like domain of Cezanne (UBACez), which lacks conserved UBA domain properties. We show that UBACez binds Lys11-, Lys48-, Lys63-, and Met1-linked ubiquitin chains in vitro, establishing UBACez as a functional ubiquitin-binding domain. Our findings also reveal that the interaction of UBACez with ubiquitin is mediated via a noncanonical surface and that hydroxylation of Asn35 inhibits ubiquitin binding. Recently, it has been suggested that Cezanne recruitment to specific target proteins depends on UBACez. Our results indicate that UBACez can indeed fulfill this role as regulatory domain by binding various ubiquitin chain types. They also uncover that this interaction with ubiquitin, and thus with modified substrates, can be modulated by oxygen-dependent asparagine hydroxylation, suggesting that Cezanne is regulated by oxygen levels.
ISSN:0021-9258
1083-351X
DOI:10.1074/jbc.RA119.010315