TY  - JOUR
AU  - Louis, Marcell
AU  - Hofmann, Kay
AU  - Broemer, Meike
TI  - Evolutionary Loss of Activity in De-Ubiquitylating Enzymes of the OTU Family.
JO  - PLOS ONE
VL  - 10
IS  - 11
SN  - 1932-6203
CY  - San Francisco, California, US
PB  - PLOS
M1  - DZNE-2020-04556
SP  - e0143227
PY  - 2015
AB  - Understanding function and specificity of de-ubiquitylating enzymes (DUBs) is a major goal of current research, since DUBs are key regulators of ubiquitylation events and have been shown to be mutated in human diseases. Most DUBs are cysteine proteases, relying on a catalytic triad of cysteine, histidine and aspartate to cleave the isopeptide bond between two ubiquitin units in a poly-ubiquitin chain. We have discovered that the two Drosophila melanogaster homologues of human OTUD4, CG3251 and Otu, contain a serine instead of a cysteine in the catalytic OTU (ovarian tumor) domain. DUBs that are serine proteases instead of cysteine- or metallo-proteases have not been described. In line with this, neither CG3251 nor Otu protein were active to cleave ubiquitin chains. Re-introduction of a cysteine in the catalytic center did not render the enzymes active, indicating that further critical features for ubiquitin binding or cleavage have been lost in these proteins. Sequence analysis of OTUD4 homologues from various other species showed that within this OTU subfamily, loss of the catalytic cysteine has occurred frequently in presumably independent events, as well as gene duplications or triplications, suggesting DUB-independent functions of OTUD4 proteins. Using an in vivo RNAi approach, we show that CG3251 might function in the regulation of Inhibitor of Apoptosis (IAP)-antagonist-induced apoptosis, presumably in a DUB-independent manner.
KW  - Amino Acid Sequence
KW  - Amino Acid Substitution
KW  - Animals
KW  - Apoptosis: genetics
KW  - Binding Sites
KW  - Catalytic Domain
KW  - Conserved Sequence
KW  - Cysteine: metabolism
KW  - Drosophila Proteins: genetics
KW  - Drosophila Proteins: metabolism
KW  - Drosophila melanogaster: genetics
KW  - Drosophila melanogaster: metabolism
KW  - Evolution, Molecular
KW  - Gene Expression Regulation
KW  - Humans
KW  - Inhibitor of Apoptosis Proteins: antagonists & inhibitors
KW  - Inhibitor of Apoptosis Proteins: genetics
KW  - Inhibitor of Apoptosis Proteins: metabolism
KW  - Molecular Sequence Data
KW  - Protein Binding
KW  - RNA, Small Interfering: genetics
KW  - RNA, Small Interfering: metabolism
KW  - Sequence Alignment
KW  - Sequence Homology, Amino Acid
KW  - Serine: metabolism
KW  - Ubiquitin: genetics
KW  - Ubiquitin: metabolism
KW  - Ubiquitin-Specific Proteases: genetics
KW  - Ubiquitin-Specific Proteases: metabolism
KW  - Ubiquitination
KW  - Drosophila Proteins (NLM Chemicals)
KW  - Inhibitor of Apoptosis Proteins (NLM Chemicals)
KW  - RNA, Small Interfering (NLM Chemicals)
KW  - Ubiquitin (NLM Chemicals)
KW  - otu protein, Drosophila (NLM Chemicals)
KW  - Serine (NLM Chemicals)
KW  - OTUD4 protein, human (NLM Chemicals)
KW  - Ubiquitin-Specific Proteases (NLM Chemicals)
KW  - Cysteine (NLM Chemicals)
LB  - PUB:(DE-HGF)16
C6  - pmid:26588485
C2  - pmc:PMC4654579
DO  - DOI:10.1371/journal.pone.0143227
UR  - https://pub.dzne.de/record/138234
ER  -