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• W2770408082 abstract "Abstract A leading objective in biology is to identify the complete set of activities performed by each gene. Identification of a comprehensive set of separation... A leading objective in biology is to identify the complete set of activities that each gene performs in vivo. In this study, we have asked whether a genetic approach can provide an efficient means of achieving this goal, through the identification and analysis of a comprehensive set of separation-of-function (sof−) mutations in a gene. Toward this goal, we have subjected the Saccharomyces cerevisiae EST1 gene, which encodes a regulatory subunit of telomerase, to intensive mutagenesis (with an average coverage of one mutation for every 4.5 residues), using strategies that eliminated those mutations that disrupted protein folding/stability. The resulting set of sof− mutations defined four biochemically distinct activities for the Est1 telomerase protein: two temporally separable steps in telomerase holoenzyme assembly, a telomerase recruitment activity, and a fourth newly discovered regulatory function. Although biochemically distinct, impairment of each of these four different activities nevertheless conferred a common phenotype (critically short telomeres) comparable to that of an est1-∆ null strain. This highlights the limitations of gene deletions, even for nonessential genes; we suggest that employing a representative set of sof− mutations for each gene in future high- and low-throughput investigations will provide deeper insights into how proteins interact inside the cell." @default.
• W2770408082 created "2017-12-04" @default.
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• W2770408082 date "2018-01-01" @default.
• W2770408082 modified "2023-09-25" @default.
• W2770408082 title "Using Separation-of-Function Mutagenesis To Define the Full Spectrum of Activities Performed by the Est1 Telomerase Subunit <i>in Vivo</i>" @default.
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• W2770408082 doi "https://doi.org/10.1534/genetics.117.300145" @default.
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