Yeast cells survive in stationary phase culture by entering quiescence, which is measured by colony-forming capacity upon nutrient re-exposure. Yeast chronological lifespan (CLS) studies, employing the comprehensive collection of gene knockout strains, have correlated weakly between independent laboratories, which is hypothesized to reflect differential interaction between the deleted genes, auxotrophy, media composition, and other assay conditions influencing quiescence. This hypothesis was investigated by high-throughput quiescence profiling of the parental prototrophic strain, from which the gene deletion strain libraries were constructed, and all possible auxotrophic allele combinations in that background. Defined media resembling human cell culture media promoted long-term quiescence and was used to assess effects of glucose, ammonium sulfate, auxotrophic nutrient availability, target of rapamycin signaling, and replication stress. Frequent, high-replicate measurements of colony-forming capacity from cultures aged past 60 days provided profiles of quiescence phenomena such as gasping and hormesis. Media acidification was assayed in parallel to assess correlation. Influences of leucine, methionine, glucose, and ammonium sulfate metabolism were clarified, and a role for lysine metabolism newly characterized, while histidine and uracil perturbations had less impact. Interactions occurred between glucose, ammonium sulfate, auxotrophy, auxotrophic nutrient limitation, aeration, TOR signaling, and/or replication stress. Weak correlation existed between media acidification and maintenance of quiescence. In summary, experimental factors, uncontrolled across previous genome-wide yeast CLS studies, influence quiescence and interact extensively, revealing quiescence as a complex metabolic and developmental process that should be studied in a prototrophic context, omitting ammonium sulfate from defined media, and employing highly replicable protocols.

酵母细胞通过进入静止期而在稳定期培养物中存活，这是通过营养物重新暴露时的集落形成能力来测量的。酵母寿命寿命（CLS）研究采用全面收集的基因敲除菌株，在独立实验室之间具有微弱的相关性，推测这反映了删除基因、营养缺陷型、培养基成分和影响静止的其他测定条件之间的差异相互作用。通过对亲本原养型菌株进行高通量静止分析来研究该假设，并从中构建了基因缺失菌株文库，以及该背景下所有可能的营养缺陷型等位基因组合。类似于人类细胞培养基的成分确定培养基促进长期静止，并用于评估葡萄糖、硫酸铵、营养缺陷型营养可用性、雷帕霉素信号传导靶标和复制应激的影响。对超过 60 天的培养物的集落形成能力进行频繁、高重复的测量，提供了静止现象的概况，例如喘息和兴奋作用。平行测定培养基酸化以评估相关性。阐明了亮氨酸、蛋氨酸、葡萄糖和硫酸铵代谢的影响，并新表征了赖氨酸代谢的作用，而组氨酸和尿嘧啶扰动的影响较小。葡萄糖、硫酸铵、营养缺陷型、营养缺陷型营养限制、通气、TOR 信号传导和/或复制应激之间发生相互作用。培养基酸化与维持静止之间存在弱相关性。总之，在之前的全基因组酵母 CLS 研究中不受控制的实验因素会影响静止并广泛相互作用，揭示静止是一个复杂的代谢和发育过程，应该在原养背景下进行研究，从确定的培养基中省略硫酸铵，并采用高度可复制的协议。