Xia Hengchuan, Shao Dandan, Liu Xiaoyong, Wang Qiang, Zhou Yang, Chen Keping
Institute of Life Sciences, Jiangsu University, 301 Xuefu Rd., Zhenjiang 212013, P. R. China.
J Insect Sci. 2017 May 1;17(3). doi: 10.1093/jisesa/iex055.
Vps20-associated 1 (Vta1) positively regulates Vacuolar protein sorting 4 (Vps4) to disassemble endosomal sorting complex required for transport III (ESCRT-III) for repeated uses in multivesicular body (MVB) pathway, virus budding and other processes. Currently, these proteins have mainly been studied in yeast and mammalian cells, while identities of them in insects remain largely unknown. We previously identified BmVps4, a Vps4 homologue from Bombyx mori. Here, we report the identification of a homologue for Vta1, designated as BmVta1. The BmVta1 cDNA contains an open reading frame of 933 bp and encodes a protein of 311 amino acid residues. We cloned BmVta1, expressed it in Escherichia coli, and prepared mouse polyclonal antibodies. Like BmVps4, BmVta1 is well conserved as shown by sequence analysis. Both proteins are localized in cytoplasm as revealed by subcellular location analysis. Interestingly, as revealed by semi-quantitative reverse transcription polymerase chain reaction (sqRT-PCR), transcriptions of BmVta1 and BmVps4 are highly up-regulated during silkworm metamorphosis and embryogenesis but down-regulated during larva stages, and are of higher levels in head, silk gland and testis than in Malpighian tube, fat body and ganglion, indicating important and similar roles of them in silkworm development and in silkworm tissues and organs. However, compared to BmVps4, the transcription of BmVta1 changes less drastically during development and is of much higher levels in midgut, ovary and hemolymph, suggesting the existence of distinct requirements of them in silkworm development and in certain tissues and organs.
Vps20相关蛋白1(Vta1)正向调节液泡蛋白分选蛋白4(Vps4),以拆解运输所需的内体分选复合物III(ESCRT-III),以便在多泡体(MVB)途径、病毒出芽及其他过程中重复使用。目前,这些蛋白主要在酵母和哺乳动物细胞中得到研究,而它们在昆虫中的特性仍 largely未知。我们之前鉴定了家蚕中的Vps4同源物BmVps4。在此,我们报告了Vta1同源物的鉴定结果,将其命名为BmVta1。BmVta1 cDNA包含一个933 bp的开放阅读框,编码一个由311个氨基酸残基组成的蛋白质。我们克隆了BmVta1,在大肠杆菌中表达,并制备了小鼠多克隆抗体。序列分析表明,与BmVps4一样,BmVta1具有高度保守性。亚细胞定位分析显示,这两种蛋白均定位于细胞质中。有趣的是,半定量逆转录聚合酶链反应(sqRT-PCR)结果表明,BmVta1和BmVps4的转录在蚕变态发育和胚胎发育过程中高度上调,但在幼虫阶段下调,且在头部、丝腺和睾丸中的水平高于马氏管、脂肪体和神经节,表明它们在蚕的发育以及蚕的组织和器官中发挥着重要且相似的作用。然而,与BmVps4相比,BmVta1的转录在发育过程中的变化较小,且在中肠、卵巢和血淋巴中的水平要高得多,这表明它们在蚕的发育以及某些组织和器官中存在不同的需求。