β1 Tubulin Rather Than β2 Tubulin Is the Preferred Binding Target for Carbendazim in Fusarium graminearum
β1 Tubulin Rather Than β2 Tubulin Is the Preferred Binding Target for Carbendazim in Fusarium graminearum
复制标题
DOI：
10.1094/phyto-09-15-0235-r
复制
发表时间：
2016-09-01
期刊：
影响因子：
3.2
通讯作者：
Zhou, Mingguo
中科院分区：
文献类型：
--
作者：
Zhou, Yujun;Zhu, Yuanye;Zhou, Mingguo
微管蛋白是抗癌药物、驱虫剂和杀真菌剂的建议靶点。在禾谷镰刀菌中，β（2）微管蛋白已被报道是苯并咪唑氨基甲酸甲酯（MBC）杀真菌剂的结合靶标。但F.与β（2）微管蛋白共享76%氨基酸序列同一性的禾谷孢菌β（1）微管蛋白在MBC敏感性中的作用尚不清楚。在这项研究中，MBC敏感性相对于亲本菌株（2021）在β（1）微管蛋白缺失菌株中显著降低，但在β（2）微管蛋白缺失菌株中增加，这表明β（1）微管蛋白参与了禾谷镰刀菌的MBC敏感性。当菌株2021在含有低或高浓度MBC杀真菌剂多菌灵（0.5或1.4 μ g/ml）的培养基中生长时，β（1）微管蛋白的蛋白质积累水平分别降低了47%和87%，而β（2）微管蛋白的蛋白质积累水平仅分别降低了6%和24%。该结果与MBC杀真菌剂更可能破坏体内GFP-β微管蛋白融合突变体中的β（1）微管蛋白微管而不是β（2）微管蛋白微管的观察结果一致。此外，序列分析表明，微管蛋白氨基酸240的差异（β（1）中的240 L与β（2）中的240 F）可能解释了MBC结合亲和力的差异;该结果与β（2）微管蛋白中的F240 L突变大大增加禾谷镰刀菌对多菌灵的敏感性的结果一致。我们认为β（1）微管蛋白而不是β（2）微管蛋白是禾本科镰刀菌MBC杀真菌剂的首选结合靶点。
Tubulins are the proposed target of anticancer drugs, anthelminthics, and fungicides. In Fusarium graminearum, beta(2) tubulin has been reported to be the binding target of methyl benzimidazole carbamate (MBC) fungicides. However, the function of F. graminearum beta(1) tubulin, which shares 76% amino acid sequence identity with beta(2) tubulin, in MBC sensitivity has been unclear. In this study, MBC sensitivity relative to that of a parental strain (2021) was significantly reduced in a beta(1) tubulin deletion strain but increased in a beta(2) tubulin deletion strain, suggesting that beta(1) tubulin was involved in the MBC sensitivity of E graminearum. When strain 2021 was grown in a medium with a low or high concentration of the MBC fungicide carbendazim (0.5 or 1.4 mu g/ml), the protein accumulation levels were reduced by 47 and 87%, respectively, for beta(1) tubulin but only by 6 and 24%, respectively, for beta(2) tubulin. This result was consistent with observations that MBC fungicides are more likely to disrupt (beta(1) tubulin microtubules rather than beta(2) tubulin microtubules in GFP-beta tubulin fusion mutants in vivo. Furthermore, sequence analysis indicated that a difference in tubulin amino acid 240 (240L in beta(1) versus 240F in beta(2)) may explain the difference in MBC binding affinity; this result was consistent with the result that an F240L mutation in beta(2) tubulin greatly increased sensitivity to carbendazim in E graminearum. We suggest that beta(1) tubulin rather than beta(2) tubulin is the preferred binding target for MBC fungicides in F graminearunz.