Novel reporter gene expression systems for monitoring activation of the Aspergillus nidulans HOG pathway
Novel reporter gene expression systems for monitoring activation of the Aspergillus nidulans HOG pathway
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DOI：
10.1271/bbb.70131
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发表时间：
2007-07-01
影响因子：
1.6
通讯作者：
Abe, Keietsu
中科院分区：
文献类型：
--
作者：
Furukawa, Kentaro;Yoshimi, Akira;Abe, Keietsu
中性曲霉高渗透压甘油反应（AnHOG）途径参与渗透适应。我们发现，杀真菌剂氟菌腈会导致假丝裂原活化蛋白激酶（MAPK）的异常激活。在这里，我们提出了新的报告系统来监测AnHOG通路的激活。将甘油-3-磷酸脱氢酶（glycerol3 -phosphate dehydrogenase, gfdB）的启动子区域与β -葡糖苷酸酶uidA基因（GUS）融合，构建报告基因，并将其引入野生型和HogA Delta中。在野生型中，只有在高渗透压或氟菌腈处理下，GUS活性才会增加，而在hogA δ突变体中，报告基因活性几乎没有受到刺激。这些结果表明，报告基因的活性是通过HogA激活来控制的。此外，我们提出了报告系统在筛选新的抗真菌化合物方面的可能应用。
The Aspergillus nidulans high-osmolarity glycerol response (AnHOG) pathway is involved in osmoadaptation. We found that fludioxonil, a fungicide, causes improper activation of HogA mitogen-activated protein kinase (MAPK) in A. nidulans. Here we present novel reporter systems for monitoring activation of the AnHOG pathway. The promoter region of gfdB (glycerol-3-phosphate dehydrogenase), whose expression depends on the presence of HogA, was fused to a beta-glucuronidase uidA gene (GUS) to construct the reporter, which was introduced into A. nidulans wild type and hogA Delta. Increased GUS activity was detected in the wild type only when it was treated with high osmolarity or fludioxonil, while reporter activity was scarcely stimulated in the hogA Delta mutant. These results indicate that the reporter activity is controlled via HogA activation. Furthermore, we present possible applications of the reporter systems in screening new antifungal compounds.