Willins D A, Xiang X, Morris N R
Department of Pharmacology, UMDNJ-Robert Wood Johnson Medical School, Piscataway 08854-5635, USA.
Genetics. 1995 Dec;141(4):1287-98. doi: 10.1093/genetics/141.4.1287.
Microtubules and cytoplasmic dynein, a microtubule-dependent motor, are required for nuclei to move along the hyphae of filamentous fungi. Nuclear migration in Aspergillus nidulans is blocked by heat-sensitive (hs-) mutations in the nudA gene, which encodes dynein heavy chain, and the nudF gene, which encodes a G protein beta-subunit-like protein. Hs- mutations in the nudC and nudG genes also prevent nuclear migration. We have isolated extragenic suppressor mutations that reverse the hs- phenotypes caused by these mutations. Here we show that one nudF suppressor also suppresses hs- mutations in nudA, nudC, and nudG and deletions in nudA and nudF. This suppressor mutation is in the tubA alpha tubulin gene, and its characteristics suggest that it destabilizes microtubules. The mutation alters microtubule staining and confers sensitivity to cold and benomyl, two treatments that destabilize microtubules. Treatment with low concentrations of benomyl also suppresses the hs- nudA, nudC, nudF, and nudG mutations and the nudA and nudF deletions. Suppression of the hs- nudA mutation and the nudA deletion is especially interesting because these strains lack active dynein heavy chain. Together, these results suggest that microtubule destabilization allows nuclei to migrate even in the absence of cytoplasmic dynein motor function.
微管和胞质动力蛋白(一种依赖微管的马达蛋白)是细胞核沿着丝状真菌菌丝移动所必需的。构巢曲霉中的核迁移会被nudA基因(编码动力蛋白重链)和nudF基因(编码一种G蛋白β亚基样蛋白)中的热敏(hs-)突变所阻断。nudC和nudG基因中的hs-突变也会阻止核迁移。我们分离出了能逆转这些突变所导致的hs-表型的基因外抑制突变。在此我们表明,一个nudF抑制子也能抑制nudA、nudC和nudG中的hs-突变以及nudA和nudF中的缺失。这个抑制突变位于tubAα微管蛋白基因中,其特征表明它会使微管不稳定。该突变改变了微管染色,并赋予对低温和苯菌灵的敏感性,这两种处理都会使微管不稳定。用低浓度苯菌灵处理也能抑制hs- nudA、nudC、nudF和nudG突变以及nudA和nudF缺失。对hs- nudA突变和nudA缺失的抑制尤其有趣,因为这些菌株缺乏活性动力蛋白重链。总之,这些结果表明,即使在没有胞质动力蛋白马达功能的情况下,微管不稳定也能使细胞核迁移。