Figure 5.
CHN-1 Ligase Activity Regulates DAF-2 Degradation and Longevity
(A and B) CHN-1::FLAG expression rescues the short lifespan of chn-1(by155) mutant worms; catalytic inactive CHN-1(U-box)::FLAG does not.
(C) DAF-2 protein level increases during aging in analyzed daf-2 mutant alleles. Importantly, daf-2(gk390525) allele displays increased DAF-2 level already at day 1 of adulthood.
(D) Short lifespan of chn-1(by155) is suppressed by daf-2(e1368) loss-of-function mutant.
(E) Deletion of chn-1 increases level of DAF-2e1368 mutant protein, analyzed by immunoblotting of worm lysates of indicated age. (F) daf-2(gk390525) worms exhibit short lifespan.
(G) Depletion of DAF-2 by RNAi extends the short lifespan of daf-2(gk390525) allele.
(H) Deletion of chn-1 does not further increase the level of DAF-2K1614R mutant protein (daf-2(gk390525) allele), analyzed by immunoblotting of worm lysates of indicated age.
(I) Loss of chn-1 shortens lifespan of daf-2(gk390525) allele to the level of chn-1(by155) worms. See Table S1 for lifespan statistics.
See also Figures S4 and S6.