IDENTIFICATION OF A GENE FOR BETA-TUBULIN IN ASPERGILLUS-NIDULANS
IDENTIFICATION OF A GENE FOR BETA-TUBULIN IN ASPERGILLUS-NIDULANS
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DOI：
10.1016/0092-8674(78)90032-6
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发表时间：
1978-01-01
期刊：
影响因子：
64.5
通讯作者：
MORRIS, NR
中科院分区：
文献类型：
--
作者：
SHEIRNEISS, G;LAI, MH;MORRIS, NR
A.通过二维凝胶电泳、与猪脑微管蛋白的共聚合和肽图谱分析，对野生型和苯菌灵A、B和C抗性菌株中的nidulans进行了表征。4 α-微管蛋白和至少4 β-通过野生型蛋白质的二维凝胶电泳解析微管蛋白。所研究的26种benA突变体中有18种具有遗传学异常的β-微管蛋白在这些菌株中，1个或多个β-微管蛋白在SDS [十二烷基硫酸钠]凝胶维度中具有改变的等电点或改变的电泳迁移率，或量减少。α-微管蛋白正常。benA/野生型二倍体菌株的蛋白质提取物的二维凝胶证明了野生型β-benA的共表达。tubulins与benA tubulin的变体。该实验排除了翻译后修饰作为β-内酰胺酶的来源。benA突变体中的微管蛋白异常。benA必须是β的结构基因-微管蛋白影响β的各种异常benA突变体中的微管蛋白，以及不存在影响α-微管蛋白的异常。任何苯菌灵抗性突变体中的微管蛋白表明苯菌灵结合位点必须位于β-微管蛋白上。微管蛋白二聚体的亚基。A. nidulans有望不仅用于表征微管蛋白的苯菌灵结合位点的生物化学决定簇，而且用于理解微管蛋白结构和功能之间的关系。
The tubulins of A. nidulans were characterized in wild-type and ben A, B and C benomyl-resistant strains by 2-dimensional gel electrophoresis, co-polymerization with porcine brain tubulin and peptide mapping. Four .alpha.-tubulins and at least 4 .beta.-tubulins were resolved by 2-dimensional gel electrophoresis of wild-type proteins. Eighteen of 26 benA mutants studied had electrophoretically abnormal .beta.-tubulins. In these strains, 1 or more of the .beta.-tubulins had either an altered isoelectric point or an altered electrophoretic mobility in the SDS [sodium dodecyl sulfate] gel dimension, or was diminished in amount. The .alpha.-tubulins were normal. Two-dimensional gels of protein extracts of a benA/wild-type diploid strain demonstrated co-expression of the wild-type .beta.-tubulins with the variant benA tubulin. This experiment rules out post-translational modification as the source of the .beta.-tubulin abnormalities in the benA mutants. benA must be a structural gene for .beta.-tubulin. The variety of abnormalities affecting .beta.-tubulins in benA mutants, and the absence of abnormalities affecting .alpha.-tubulins in any of the benomyl-resistant mutants indicate that the benomyl binding site must be located on the .beta.-subunit of the tubulin dimer. The benA mutants of A. nidulans promise to be useful not only for characterizing the biochemical determinants of the benomyl binding site of tubulin but also for understanding the relationship between tubulin structure and function.