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Abstract
We describe a new chronological lifespan (CLS) assay for the yeast Schizosaccharomyces
pombe. Yeast CLS assays monitor the loss of cell viability in a culture over time,
and this new assay shows a continuous decline in viability without detectable regrowth
until all cells in the culture are dead. Thus, the survival curve is not altered by
the generation of mutants that can grow during the experiments, and one can monitor
the entire lifespan of a strain until the number of viable cells has decreased over
10(6)-fold. This CLS assay recapitulates the evolutionarily conserved features of
lifespan shortening by over nutrition, lifespan extension by caloric restriction,
increased stress resistance of calorically restricted cells and lifespan control by
the AKT kinases. Both S. pombe AKT kinase orthologs regulate CLS: loss of sck1(+)
extended lifespan in over nutrition conditions, loss of sck2(+) extended lifespan
under both normal and over nutrition conditions, and loss of both genes showed that
sck1(+) and sck2(+) control different longevity pathways. The longest-lived S. pombe
cells showed the most efficient cell cycle exit, demonstrating that caloric restriction
links these two processes. This new S. pombe CLS assay will provide a valuable tool
for aging research.