Low auxotrophy-complementing amino acid concentrations reduce yeast chronological life span

In the yeast Saccharomyces cerevisiae, interventions resembling caloric restriction, either by reduction of glucose or non-essential amino acid content in the medium, prolong life span and retard aging. Here we have examined the role of auxotrophy-complementing amino acid supplementation of S. cerevisiae strains in determining yeast chronological life span and stress resistance. The results obtained from cells cultured in standard amino acid concentrations revealed a reduced final biomass yield and premature aging phenotypes. These included shorter life span and indicators of oxidative stress, together with a G2/M cell cycle arrest and the appearance of a sub-G0/G1 population pointing to the occurrence of a specific cell death programme under starvation of essential amino acids. In order to overcome this starvation, five times higher amino acid concentrations were supplied to the medium as has already been commonly used by few laboratories. Such cultures reached more than five-fold higher final biomass yield in stationary phase and the early aging phenotypes were abrogated. Furthermore, in a long-lived yeast strain lacking TOR1, there was no positive effect of amino acid supplementation on longevity. On the contrary, amino acid supply had a positive effect on chronological life span of RAS2 deleted cells. This study may provide novel insights into the role of essential nutrients and their effect on aging process and raises the warning that the positive effects of caloric restriction on life span maybe restricted to non-essential nutrients. Moreover, the severe consequences on cell physiology, life span and stress resistance induced by essential amino acid imbalances presents a note of caution for those still using standard amino acid concentrations for studies with auxotrophic yeast strains.We thank members of the Ludovico and Rodrigues labs forassistance with figures, helpful discussions and comments.Pedro Gomes and Belém Sampaio-Marques were financiallysupported by fellowships from Fundação para a Ciência e Tecnologia, Portugal (SFRH/BPD/13969/2003 and SFRH/BI/15406/2005). This work was supported by a grant awarded byFundação para a Ciência e Tecnologia (POCI/BIA-BCM/57364/2004