HEAT-INDUCIBLE DEGRON - A METHOD FOR CONSTRUCTING TEMPERATURE-SENSITIVE MUTANTS
HEAT-INDUCIBLE DEGRON - A METHOD FOR CONSTRUCTING TEMPERATURE-SENSITIVE MUTANTS
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DOI：
10.1126/science.8122109
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发表时间：
1994-03-04
期刊：
影响因子：
56.9
通讯作者：
VARSHAVSKY, A
中科院分区：
文献类型：
--
作者：
DOHMEN, RJ;WU, PP;VARSHAVSKY, A
温度敏感突变体在低温（允许）下保留基因的功能，但在高温（不允许）下则不保留。Arg-DHFR是一种带有氨基末端（N末端）精氨酸的二氢叶酸还原酶，在酵母酿酒酵母中寿命很长，尽管精氨酸是蛋白质降解N末端规则中的不稳定残基。鉴定了Arg-DHFR的ts衍生物，其在23 ℃下长寿命，但在37 ℃下通过N-末端规则途径快速降解。将ts Arg-DHFR与S.酿酒酵母赋予这些基因产物特异性的TS表型。因此，Arg-DHFR（ts）是热诱导降解信号，其可用于产生ts突变体而无需寻找ts突变。
A temperature-sensitive (ts) mutant retains the function of a gene at a low (permissive) temperature but not at a high (nonpermissive) temperature. Arg-DHFR, a dihydrofolate reductase bearing an amino-terminal (N-terminal) arginine, is long-lived in the yeast Saccharomyces cerevisiae, even though arginine is a destabilizing residue in the N-end rule of protein degradation. A ts derivative of Arg-DHFR was identified that is long-lived at 23 degrees C but rapidly degraded by the N-end rule pathway at 37 degrees C. Fusions of ts Arg-DHFR to either Ura3 or Cdc28 of S. cerevisiae confer ts phenotypes specific for these gene products. Thus, Arg-DHFR(ts) is a heat-inducible degradation signal that can be used to produce ts mutants without a search for ts mutations.