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TRIP12 ubiquitinates OTUD5. 11 / 11
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"TRIP12 promotes OTUD5 ubiquitylation and degradation."
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"We subsequently investigated whether TRIP12 promoted ubiquitylation of OTUD5 in cell and in vitro."
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"Using TUBE2 (tandem ubiquitin-binding entity 2), which serves as a pan-ubiquitin chain binder when four repeats of the UBA1 domains from HR23A are tandemly fused , we found that OTUD5 pan-ubiquitylation was enhanced by the expression of wild-type, but not catalytically inactive (C1959A), TRIP12 (Fig. 1f)."
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"We also confirmed that TRIP12 knockdown decreased ubiquitylation of endogenous OTUD5 under proteasomal inhibition (Fig. 1g)."
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"In an in vitro ubiquitylation assay using purified OTUD5 and TRIP12 proteins (supplementary Fig. 1c), OTUD5 was ubiquitylated by TRIP12 (Fig. 1h)."
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"However, wild-type and C224R OTUD5 were ubiquitylated by TRIP12 at similar levels (Fig.  xref (ii))."
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"The data on a catalytically-inactive OTUD5 (Fig.  xref ) suggest that the ubiquitylation of OTUD5 by TRIP12 and UBR5 appears as if they act sequentially because OTUD5 DUB activity negates UBR5 activity, which is different from typical chain initiation (priming)-elongation model xref , xref ."
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"UBR5 expression promoted both K29- and K48-linked ubiquitylation of OTUD5, which was almost completely diminished by TRIP12 knockdown (Fig. 2i, lanes 2, 4, 6, and 8)."
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"The data on a catalytically-inactive OTUD5 (Fig. 5) suggest that the ubiquitylation of OTUD5 by TRIP12 and UBR5 appears as if they act sequentially because OTUD5 DUB activity negates UBR5 activity, which is different from typical chain initiation (priming)-elongation model ."
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"Next, we investigated whether OTUD5 DUB activity counteracted OTUD5 ubiquitylation by TRIP12 or UBR5."
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"In an in vitro ubiquitylation assay using purified OTUD5 and TRIP12 proteins (supplementary Fig.  xref ), OTUD5 was ubiquitylated by TRIP12 (Fig.  xref )."
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