Department of Molecular Genetics, Cleveland Clinic Lerner College of Medicine at Case Western Reserve University, Cleveland, Ohio, USA.
PLoS One. 2013 Jul 9;8(7):e69084. doi: 10.1371/journal.pone.0069084. Print 2013.
Model organisms such as budding yeast, worms and flies have proven instrumental in the discovery of genetic determinants of aging, and the fission yeast Schizosaccharomyces pombe is a promising new system for these studies. We devised an approach to directly select for long-lived S. pombe mutants from a random DNA insertion library. Each insertion mutation bears a unique sequence tag called a bar code that allows one to determine the proportion of an individual mutant in a culture containing thousands of different mutants. Aging these mutants in culture allowed identification of a long-lived mutant bearing an insertion mutation in the cyclin gene clg1(+). Clg1p, like Pas1p, physically associates with the cyclin-dependent kinase Pef1p. We identified a third Pef1p cyclin, Psl1p, and found that only loss of Clg1p or Pef1p extended lifespan. Genetic and co-immunoprecipitation results indicate that Pef1p controls lifespan through the downstream protein kinase Cek1p. While Pef1p is conserved as Pho85p in Saccharomyces cerevisiae, and as cdk5 in humans, genome-wide searches for lifespan regulators in S. cerevisiae have never identified Pho85p. Thus, the S. pombe system can be used to identify novel, evolutionarily conserved lifespan extending mutations, and our results suggest a potential role for mammalian cdk5 as a lifespan regulator.
模式生物,如芽殖酵母、线虫和果蝇,已被证明在发现衰老的遗传决定因素方面具有重要作用,裂殖酵母 Schizosaccharomyces pombe 是这些研究的一种很有前途的新系统。我们设计了一种方法,从随机 DNA 插入文库中直接筛选长寿 S. pombe 突变体。每个插入突变都带有一个独特的序列标签,称为条形码,允许确定含有数千种不同突变体的培养物中单个突变体的比例。在培养物中使这些突变体衰老,可鉴定出一个携带 cyclin 基因 clg1(+)插入突变的长寿突变体。Clg1p 与 Pas1p 一样,与细胞周期蛋白依赖性激酶 Pef1p 物理结合。我们鉴定了第三种 Pef1p 细胞周期蛋白 Psl1p,并发现只有 Clg1p 或 Pef1p 的缺失才能延长寿命。遗传和共免疫沉淀结果表明,Pef1p 通过下游蛋白激酶 Cek1p 控制寿命。虽然 Pef1p 在酿酒酵母中与 Pho85p 保守,在人类中与 cdk5 保守,但在酿酒酵母中进行的全基因组搜索从未发现过 Pho85p 作为寿命调节剂。因此,S. pombe 系统可用于鉴定新的、进化上保守的延长寿命的突变体,我们的结果表明哺乳动物 cdk5 可能作为寿命调节剂发挥作用。