Figure 4.

Prefoldin-like complex components are required for the long life span of daf-2 mutants. (A) Schematic model of the R2TP/prefoldin-like complex and the prefoldin complex in mammals; the R2TP/prefoldin-like complex interacts with RNA polymerase II in the cytoplasm to help its assembly, and the prefoldin complex is composed of PFD-1, PFD-2, PFD-3, PFD-4, PFD-5, and PFD-6, which bind to nascent actin or tubulin polypeptides. Prefoldin complex-bound polypeptides are transferred to TRiC/CCT chaperonin for proper folding. (B) Yeast two-hybrid screening with full-length PFD-6 as bait was performed using the yeast PBN204 strain. The interaction was validated using nutritional selection (ADE2) for the growth assay. See Supplemental Table S7 for the description of candidate genes. (C–F) Several R2TP/prefoldin-like complex components were required for life span extension caused by reduced IIS. RNAi targeting ruvb-1 (C), ruvb-2 (D), uri-1 (E), or pfd-2 (F) significantly decreased the long life span of daf-2(e1370) [daf-2(−)] mutants. Knockdown of potential UXT homolog F35H10.6 did not affect the life span of wild-type, daf-2(−), pfd-6(−), or pfd-6; daf-2 mutants (Supplemental Fig. S4C,G). See Supplemental Table S5 for statistical analysis and additional repeats.