Investigation of Isoform Specific Functions of the V-ATPase a Subunit During Drosophila Wing Development
Investigation of Isoform Specific Functions of the V-ATPase a Subunit During Drosophila Wing Development
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果蝇翅膀发育过程中 V-ATP 酶 a 亚基异构体特异性功能的研究
DOI：
10.3389/fgene.2020.00723
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发表时间：
2020-07-10
影响因子：
3.7
通讯作者：
Zhang, Junzheng
中科院分区：
文献类型：
--
作者：
Mo, Dongqing;Chen, Yao;Zhang, Junzheng
液泡ATP酶（V-ATP酶）是ATP依赖的质子泵，在真核细胞中起着重要作用。昆虫V-ATP酶在几乎所有的上皮组织中都是必需的，以调节多种过程，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，几个V-ATP酶亚基以不同的同种型存在，以执行细胞类型特异性功能。V-ATP酶的100 kD α亚基（Vha 100）由果蝇中的一个5基因家族编码，但它们的归属还不完全清楚。本文报道了果蝇翅发育过程中Vha 100基因家族的实验研究。使用CRISPR-Cas9诱变、体细胞克隆分析和体内RNAi测定的组合来表征Vha 100同种型的需求，并产生Vha 100 -2、Vha 100 -3、Vha 100 -4和Vha 100 - 5基因的突变体。结果表明，Vha 100 - 3和Vha 100 - 5对果蝇的发育是有利的，而Vha 100 - 1对果蝇的发育不是必需的。另外两种亚型中，Vha 100 - 2参与调控翅表皮成熟，而Vha 100 - 4是参与发育模式形成的唯一亚型。更具体地说，Vha 100 - 4是在苍蝇翅膀发育过程中正确激活无翅信号通路所必需的。有趣的是，我们还发现Vha 100 - 1和Vha 100 - 4在翅膀发育过程中存在特异的遗传互作。我们的研究结果揭示了Vha 100亚型在昆虫翅膀发育过程中的不同作用，为理解V-ATP酶的不同作用提供了理论基础。
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 inDrosophila, but their assignments are not fully understood. Here we report an experimental survey of theVha100gene family duringDrosophilawing development. A combination of CRISPR-Cas9 mutagenesis, somatic clonal analysis andin vivoRNAi assays is used to characterize the requirement of Vha100 isoforms, and mutants ofVha100-2,Vha100-3, Vha100-4, andVha100-5genes were generated. We show thatVha100-3andVha100-5are dispensable for fly development, whileVha100-1is not critically required in the wing. As for the other two isoforms, we find thatVha100-2regulates wing cuticle maturation, whileVha100-4is the single isoform involved in developmental patterning. More specifically,Vha100-4is required for proper activation of the Wingless signaling pathway during fly wing development. Interestingly, we also find a specific genetic interaction betweenVha100-1andVha100-4during wing development. Our results revealed the distinct roles ofVha100isoforms during insect wing development, providing a rationale for understanding the diverse roles of V-ATPases.