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Biochemical assays In vitro ubiquitylation assays The identification of cullin–RING ligase subunits and the ARIH1 RBR-type E3 as TDMD effectors 10 suggested that ZSWIM8 performs ubiquitylation by the previously defined E3–E3 mechanism, which is a variation on the canonical E1–E2–E3 pathway 35,36
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