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Investigation of Isoform Specific Functions of the V-ATPase a Subunit During Drosophila Wing Development.
Frontiers in Genetics ( IF 3.7 ) Pub Date : 2020-06-15 , DOI: 10.3389/fgene.2020.00723
Dongqing Mo 1 , Yao Chen 1 , Na Jiang 1 , Jie Shen 1 , Junzheng Zhang 1
Affiliation  

The vacuolar ATPases (V-ATPases) are ATP-dependent proton pumps that play vital roles in eukaryotic cells. Insect V-ATPases are required in nearly all epithelial tissues to regulate a multiplicity of processes including receptor-mediated endocytosis, protein degradation, fluid secretion, and neurotransmission. Composed of fourteen different subunits, several V-ATPase subunits exist in distinct isoforms to perform cell type specific functions. The 100 kD a subunit (Vha100) of V-ATPases are encoded by a family of five genes in Drosophila, but their assignments are not fully understood. Here we report an experimental survey of the Vha100 gene family during Drosophila wing development. A combination of CRISPR-Cas9 mutagenesis, somatic clonal analysis and in vivo RNAi assays is used to characterize the requirement of Vha100 isoforms, and mutants of Vha100-2, Vha100-3, Vha100-4, and Vha100-5 genes were generated. We show that Vha100-3 and Vha100-5 are dispensable for fly development, while Vha100-1 is not critically required in the wing. As for the other two isoforms, we find that Vha100-2 regulates wing cuticle maturation, while Vha100-4 is the single isoform involved in developmental patterning. More specifically, Vha100-4 is required for proper activation of the Wingless signaling pathway during fly wing development. Interestingly, we also find a specific genetic interaction between Vha100-1 and Vha100-4 during wing development. Our results revealed the distinct roles of Vha100 isoforms during insect wing development, providing a rationale for understanding the diverse roles of V-ATPases.



中文翻译:

果蝇翼发育过程中V-ATPase亚基的同工型特异性功能的研究。

液泡ATPase(V-ATPase)是依赖ATP的质子泵,在真核细胞中起重要作用。几乎所有上皮组织都需要昆虫V-ATPase来调节多种过程,包括受体介导的内吞作用,蛋白质降解,液体分泌和神经传递。由14个不同的亚基组成的多个V-ATPase亚基以不同的同工型存在,以执行细胞类型特异性功能。V-ATPase的100 kD a亚基(Vha100)由5个基因的家族编码果蝇,但尚未完全了解其分配。在这里,我们报告了对Vha100 期间的基因家族 果蝇机翼的发展。CRISPR-Cas9诱变,体细胞克隆分析和体内 RNAi分析用于表征Vha100同工型和 Vha100-2Vha100-3,Vha100-4Vha100-5基因产生。我们证明Vha100-3Vha100-5 对于苍蝇发育是必不可少的 Vha100-1在机翼中不是十分必要的。至于其他两个同工型,我们发现Vha100-2 调节机翼角质层的成熟,而 Vha100-4是参与发育模式的单一同工型。进一步来说,Vha100-4在机翼发育过程中正确激活无翼信号通路是必需的。有趣的是,我们还发现Vha100-1Vha100-4在机翼开发过程中。我们的结果揭示了Vha100 昆虫翅膀发育过程中的同工型,为理解V-ATPase的不同作用提供了理论依据。

更新日期:2020-07-10
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