Crosstalk Between Pheromone Signaling and NADPH Oxidase Complexes Coordinates Fungal Developmental Processes
Crosstalk Between Pheromone Signaling and NADPH Oxidase Complexes Coordinates Fungal Developmental Processes
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DOI：
10.3389/fmicb.2020.01722
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发表时间：
2020-07
影响因子：
5.2
通讯作者：
Sarah Schmidt;R. Märker;Barbara Ramšak;Anna M. Beier-Rosberger;I. Teichert;U. Kück
中科院分区：
文献类型：
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作者：
Sarah Schmidt;R. Märker;Barbara Ramšak;Anna M. Beier-Rosberger;I. Teichert;U. Kück
丝状子囊菌的有性和无性发育是由保守的信号通路控制的。在这里，我们研究了缺乏激酶MAK2，MEK2和MIK2基因的突变株的发展，以及信息素反应（PR）途径的支架蛋白HAM5。结果表明，所有菌株子实体发育和菌丝融合均存在缺陷。另一种表型是黑色素依赖的子囊孢子萌发缺陷。然而，这种缺陷仅在激酶缺失突变体中观察到，而在缺乏HAM5的菌株中未观察到。值得注意的是，先前描述了烟酰胺腺嘌呤二核苷酸磷酸（NADPH）氧化酶1（NOX 1）突变体的相同发育表型，但发芽缺陷仅见于NOX 2突变体。这些数据表明信息素信号通路和两个NOX复合物之间的分子联系。利用酵母双杂交（Y2H）分析的数据，我们发现支架蛋白HAM5与NOR1相互作用，NOX1和NOX2复合物的调节剂。使用不同的荧光标记蛋白进一步证实了这种相互作用，以证明NOR1和HAM5共定位于成熟菌丝的细胞质点和尖端。单突变体和双突变体的表型表征支持这一观察结果。Δham5、Δnor1和Δham5的氧化应激反应和子实体的起始相似，但Δnor1的氧化应激反应和子实体的起始明显降低，表明双缺失导致Δnor1表型的部分抑制。我们的结论是PR和NOX 1复合物连接HAM5和NOR1之间的直接相互作用。相比之下，PR激酶与N0X2复合物连接而不参与HAM5。
Sexual and asexual development in filamentous ascomycetes is controlled by components of conserved signaling pathways. Here, we investigated the development of mutant strains lacking genes for kinases MAK2, MEK2, and MIK2, as well as the scaffold protein HAM5 of the pheromone response (PR) pathway. All had a defect in fruiting body development and hyphal fusion. Another phenotype was a defect in melanin-dependent ascospore germination. However, this deficiency was observed only in kinase deletion mutants, but not in strains lacking HAM5. Notably, the same developmental phenotypes were previously described for nicotinamide adenine dinucleotide phosphate (NADPH) oxidase 1 (NOX1) mutants, but the germination defect was only seen in NOX2 mutants. These data suggest a molecular link between the pheromone signaling pathway and both NOX complexes. Using data from yeast two-hybrid (Y2H) analysis, we found that the scaffolding protein HAM5 interacts with NOR1, the regulator of NOX1 and NOX2 complexes. This interaction was further confirmed using differently fluorescent-labeled proteins to demonstrate that NOR1 and HAM5 co-localize at cytoplasmic spots and tips of mature hyphae. This observation was supported by phenotypic characterization of single and double mutants. The oxidative stress response and the initiation of fruiting bodies were similar in Δham5Δnor1 and Δham5, but distinctly reduced in Δnor1, indicating that the double deletion leads to a partial suppression of the Δnor1 phenotype. We conclude that the PR and NOX1 complexes are connected by direct interaction between HAM5 and NOR1. In contrast, PR kinases are linked to the NOX2 complex without participation of HAM5.