a microarray-based genetic screen for yeast chronological aging factors基因芯片筛选酵母老化时间因素.pdfVIP

A Microarray-Based Genetic Screen for Yeast Chronological Aging Factors 1 1 2 1 1 2 Mirela Matecic , Daniel L. Smith, Jr. , Xuewen Pan , Nazif Maqani , Stefan Bekiranov , Jef D. Boeke , Jeffrey S. Smith1* 1 Department of Biochemistry and Molecular Genetics, University of Virginia Health System, School of Medicine, Charlottesville, Virginia, United States of America, 2 Department of Molecular Biology and Genetics, High Throughput Biology Center, Johns Hopkins University School of Medicine, Baltimore, Maryland, United States of America Abstract Model organisms have played an important role in the elucidation of multiple genes and cellular processes that regulate aging. In this study we utilized the budding yeast, Saccharomyces cerevisiae, in a large-scale screen for genes that function in the regulation of chronological lifespan, which is defined by the number of days that non-dividing cells remain viable. A pooled collection of viable haploid gene deletion mutants, each tagged with unique identifying DNA ‘‘bar-code’’ sequences was chronologically aged in liquid culture. Viable mutants in the aging population were selected at several time points and then detected using a microarray DNA hybridization technique that quantifies abundance of the barcode tags. Multiple short- and long-lived mutants were identified using this approach. Among the confirmed short-lived mutants were those defective for autophagy, indicating a key requirement for the recycling of cellular organelles in longevity. Defects in autophagy also prevented lifespan extension induced by limitation of amino acids in the growth media. Among the confirmed long-lived mutants were those defective in the highly conserved de