Mutant Caenorhabditis elegans RNA polymerase II with a 20,000-fold reduced sensitivity to alpha-amanitin.
Mutant Caenorhabditis elegans RNA polymerase II with a 20,000-fold reduced sensitivity to alpha-amanitin.
复制标题
突变的秀丽隐杆线虫 RNA 聚合酶 II 对 α-鹅膏蕈碱的敏感性降低了 20,000 倍。
DOI：
10.1093/genetics/126.4.889
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发表时间：
1990
期刊：
影响因子：
3.3
通讯作者：
Riddle,DL
中科院分区：
文献类型：
--
作者：
Rogalski,TM;Golomb,M;Riddle,DL
一个双重突变的ama-1（m118 m526）基因导致RNA聚合酶（Rpo）II对α-鹅膏蕈碱具有不寻常的抗性。在分离的秀丽隐杆线虫细胞核中的Rpo II活性被浓度为150微克/毫升的α-鹅膏蕈碱抑制50%，使得该酶对毒素的抗性比来自单突变等位基因ama-1（m118）的Rpo II高150倍，比野生型Rpo II高20，000倍，并且对鹅膏蕈碱的抗性比Rpo III高约6倍。确定SL 1剪接前导前体由Rpo II转录，并且该转录物用于测量Rpo II活性。Rpo II活性在体外不稳定，突变株具有温度敏感的不育表型。在诱变ama-1（m118）亲本的400万后代中，通过其在透化剂Triton X-100存在下在200微克/ml鹅膏蕈碱中生长和繁殖的能力，选择了高度抗性的双突变体。
A doubly mutant ama-1(m118m526) gene results in an RNA polymerase (Rpo) II that is unusually resistant to alpha-amanitin. Rpo II activity in isolated Caenorhabditis elegans cell nuclei is inhibited 50% by alpha-amanitin at a concentration of 150 micrograms/ml, making this enzyme 150 times more resistant to the toxin than Rpo II from the singly mutant allele, ama-1(m118), 20,000 times more resistant than the wild-type Rpo II, and about six times more resistant to amanitin than is Rpo III. It was determined that the SL1 spliced leader precursor is transcribed by Rpo II, and this transcript was used to measure Rpo II activity. The Rpo II activity is unstable in vitro, and the mutant strain has a temperature-sensitive sterile phenotype. The highly resistant double mutant was selected among four million progeny of the mutagenized ama-1(m118) parent by its ability to grow and reproduce in 200 micrograms/ml amanitin in the presence of a permeabilizing agent, Triton X-100.