Table 1.
ATG13 Is Phosphorylated at Known Repressive Sites during Mitosis
| Treatment (Fold Change to DMSO Control) |
||||||
|---|---|---|---|---|---|---|
| Paclitaxel | AZD8055 | Paclitaxel + AZD8055 | ||||
| Phosphopeptide phosphorylation site and relevant interphase kinase | Rep 1 | Rep 2 | Rep 1 | Rep 2 | Rep 1 | Rep 2 |
| TPPIMGIIIDHFVDRPYPSSSPMHPCNYR S224 (known AMPK-dependent site) | 3.55 | 2.07 | 0.94 | 0.96 | 3.86 | 3.00 |
| TAGEDTGVIYPSVEDSQEVCTTSFSTSPPSQLSSSR S259 (known mTOR site) | 1.75 | 1.90 | 0.15 | 0.19 | 0.81 | 1.05 |
HEK293 GFP-ATG13 cells were treated with paclitaxel (50 nM, 16 h) and/or AZD8055 (1 μM, 2 h). GFP-ATG13 was then immunoprecipitated, protease digested, and analyzed by liquid chromatography tandem mass spectrometry as outlined in STAR Methods. Data for two phosphopeptides are shown; during interphase, S224 is known to be phosphorylated in an AMPK-dependent manner and S259 by mTOR directly. Fold-change compared to DMSO control is presented for two independent replicate experiments.