‘Evolve and resequence’ (E&R) studies in
Drosophila melanogaster
have identified many candidate loci underlying the evolution of ageing and life history, but experiments that validate the effects of such candidates remain rare. In a recent E&R study we have identified several alleles of the LAMMER kinase
Darkener of apricot
(
Doa
) as candidates for evolutionary changes in lifespan and fecundity. Here, we use two complementary approaches to confirm a functional role of
Doa
in life-history evolution. First, we used transgenic RNAi to study the effects of
Doa
at the whole-gene level. Ubiquitous silencing of expression in adult flies reduced both lifespan and fecundity, indicating pleiotropic effects. Second, to characterize segregating variation at
Doa
, we examined four candidate single nucleotide polymorphisms (SNPs;
Doa-1
, -
2
, -
3
,
-4
) using a genetic association approach. Three candidate SNPs had effects that were qualitatively consistent with expectations based on our E&R study:
Doa-2
pleiotropically affected both lifespan and late-life fecundity;
Doa-1
affected lifespan (but not fecundity); and
Doa-4
affected late-life fecundity (but not lifespan). Finally, the last candidate allele (
Doa-3
) also affected lifespan, but in the opposite direction from predicted.