Extended Data Fig. 9: WDR24-WDR59 α-solenoid junction is required for GATOR2 assembly and function.

(a) Two views of the WDR24-WDR59 α-solenoid junction. (b) Mutations in the WDR24 α-solenoid domain disrupt the WDR24-WDR59 interaction. Anti-FLAG immunoprecipitates were prepared from MIOS-deficient HEK293T cells that transiently expressed the indicated cDNAs and were analyzed by immunoblotting for the indicated proteins. HA, hemagglutinin. (c) The WDR24-WDR59 interaction is required for activation of mTORC1 and the GATOR2-GATOR1 and GATOR2-KICSTOR interactions. Anti-FLAG and anti-HA immunoprecipitates were prepared from WDR24-deficient HEK293T cells that transiently expressed the indicated cDNAs and were analyzed as in (b). (d) Mutation of the WDR59 solenoidal domain disrupts the WDR24-WDR59 interaction. Anti-FLAG immunoprecipitates were prepared from MIOS-deficient HEK293T cells that transiently expressed the indicated cDNAs and were analyzed as in (b). (e) The WDR24-WDR59 interaction is required for activation of mTORC1 and the GATOR2-GATOR1 and GATOR2-KICSTOR interactions. Anti-FLAG and anti-HA immunoprecipitates were prepared from WDR24-deficient HEK293T cells that transiently expressed the indicated cDNAs and were analyzed as in (b). Data in (b)–(e) are representative of two independent experiments. For gel source data, see Supplementary Figure 1.