Cabrera Ilva E, Oza Yagna, Carrillo Alexander J, Collier Logan A, Wright Sara J, Li Liande, Borkovich Katherine A
Department of Microbiology and Plant Pathology, University of California-Riverside, 900 University Avenue, Riverside, CA 92521, USA.
J Fungi (Basel). 2022 Oct 13;8(10):1076. doi: 10.3390/jof8101076.
Heterotrimeric (αβγ) G protein signaling pathways are critical environmental sensing systems found in eukaryotic cells. Exchange of GDP for GTP on the Gα subunit leads to its activation. In contrast, GTP hydrolysis on the Gα is accelerated by Regulator of G protein Signaling (RGS) proteins, resulting in a return to the GDP-bound, inactive state. Here, we analyzed growth, development and extracellular cellulase production in strains with knockout mutations in the seven identified RGS genes ( to ) in the filamentous fungus, We compared phenotypes to those of strains with either knockout mutations or expressing predicted constitutively activated, GTPase-deficient alleles for each of the three Gα subunit genes (, or ). Our data revealed that six RGS mutants have taller aerial hyphae than wild type and all seven mutants exhibit reduced asexual sporulation, phenotypes shared with strains expressing the or allele. In contrast, Δ and Δ were the only RGS mutants with a slower growth rate phenotype, a defect in common with strains. With respect to female sexual development, Δ possessed defects most similar to strains, while those of Δ mutants resembled strains expressing the allele. Finally, we observed that four of the seven RGS mutants had significantly different extracellular cellulase levels relative to wild type. Of interest, the Δ mutant had no detectable activity, similar to the strain. In contrast, the Δ and Δ mutants and and strains exhibited significantly higher cellulase activity than wild type. With the exception of sexual development, our results demonstrate the greatest number of genetic interactions between and and and in .
异源三聚体(αβγ)G蛋白信号通路是真核细胞中重要的环境感知系统。Gα亚基上的GDP与GTP交换会导致其激活。相反,G蛋白信号调节(RGS)蛋白会加速Gα上的GTP水解,导致其回到结合GDP的无活性状态。在此,我们分析了丝状真菌中七个已鉴定的RGS基因(至)敲除突变体菌株的生长、发育和细胞外纤维素酶产生情况。我们将这些突变体的表型与三个Gα亚基基因(、或)的敲除突变体或表达预测的组成型激活、GTP酶缺陷等位基因的菌株的表型进行了比较。我们的数据显示,六个RGS突变体的气生菌丝比野生型更高,所有七个突变体的无性孢子形成均减少,这些表型与表达或等位基因的菌株相同。相比之下,Δ和Δ是仅有的具有生长速率较慢表型的RGS突变体,这一缺陷与菌株相同。关于雌性有性发育,Δ具有与菌株最相似的缺陷,而Δ突变体的缺陷类似于表达等位基因的菌株。最后,我们观察到七个RGS突变体中有四个相对于野生型具有显著不同的细胞外纤维素酶水平。有趣的是,Δ突变体没有可检测到的活性,类似于菌株。相比之下,Δ和Δ突变体以及和菌株表现出比野生型显著更高的纤维素酶活性。除有性发育外,我们的结果表明在中与和与之间存在最多的遗传相互作用。