Figure 5

Epistasis testing of kinases apparently required to prevent protein degradation against known pathways. (A) The 48 kinases that were identified as required to prevent protein degradation (Figure 1B) were examined for potential interaction with known proteolytic signalling pathways in C. elegans muscle [22, 34, 39]. For these experiments the acute RNAi screen, see materials and methods, was rerun on a control set of PD55 animals (not shown) and on PJ1009 (unc-51(e369), which has been shown to block autophagic degradation [22]), PD55 treated with MG132 (which has been shown to block proteasomal degradation [39]), PJ1103 (mpk-1(n2521), which has been shown to block degradation resulting from excessive FGFR or insufficient IGFR signalling [22]), and PJ1132 (daf-18(e1375), which has been shown to block degradation resulting from excessive FGFR or insufficient IGFR signalling[22]). At least two independent experiments per gene per strain were performed, with a third and/or more run in case of discrepant results. (B) Kinase knockdowns identified as requiring autophagy, the proteasome, or another proteolytic system to induce degradation are indicated by the columns on the left. Kinase knockdowns identified as requiring MPK-1 and/or DAF-18 for muscle protein degradation are indicated by the columns on the right. The observation, or lack of observation, of protein degradation caused by knockdown of the indicated kinase (middle columns), in at least 2 experiments, in each of the test conditions is indicated by a colour code for which an inset legend is provided.