Figure 6.

Mimicking Phosphorylation of ATL1 on S312 Inhibits Its in Vitro Ubiquitination Activity.

In vitro ubiquitination assays were performed in the presence of the E1 protein Uba2, the E2 protein UbcH5a, and Mg-ATP, and ATL1 autoubiquitination was detected using an antiubiquitin antibody. Multiple ubiquitinated protein forms were detected when wild-type ATL1 was used (lane 1). No ubiquitination activity was detected using ATL1^S312D (lane 3). ATL1^R308A (lane 2) and ATL1^S312A (lane 4) retained ubiquitination activity.