Molecular cloning, expression and characterization of a novel vacuolar protein sorting 4 gene in silkworm, Bombyx mori
Molecular cloning, expression and characterization of a novel vacuolar protein sorting 4 gene in silkworm, Bombyx mori
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DOI：
10.1007/s11033-012-1911-6
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发表时间：
2012-12-01
影响因子：
2.8
通讯作者：
Chen, Keping
中科院分区：
文献类型：
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作者：
Xia, Hengchuan;Zhang, Chunxia;Chen, Keping
液泡蛋白分选4 （Vps4）蛋白在多泡体（MVB）途径、病毒出芽过程和细胞分裂过程中起重要作用。Vps4已在许多种家蚕中被鉴定和鉴定，但尚未在家蚕中发现。本研究首次鉴定并克隆了VPS4的家蚕同源基因，并在大肠杆菌中表达，纯化并鉴定了其蛋白BmVps4。BmVps4 cDNA包含1,314 bp的开放阅读框，编码438个氨基酸残基的蛋白。BmVps4与其他物种的Vps4蛋白序列相似性高。重组BmVps4具有atp酶活性，可以被Mg2+激活，并被显性突变抑制。总之，我们的数据表明BmVps4是Vps4的真正的家蚕同源物。据我们所知，这是首次对蚕MVB蛋白进行表征。该研究将为进一步研究家蚕MVB途径及其在家蚕最常见和最严重的病原体之一家蚕核型多角体病毒（BmNPV）感染和出芽过程中的可能作用提供依据。克隆的BmVps4序列存入GenBank（登录号GQ995504）。
The vacuolar protein sorting 4 (Vps4) protein is essential for the multivesicular body (MVB) pathway, virus budding process and cytokinesis. Vps4 has been identified and characterized from many species, but not from silkworm Bombyx mori. In this study, we firstly identified and cloned the silkworm homologous gene for VPS4, expressed it in Escherichia coli, purified and characterized the protein designated as BmVps4. The BmVps4 cDNA contains an open reading frame of 1,314 bp, and encodes a protein of 438 amino acid residues. BmVps4 is of high sequence-similarity to Vps4 proteins from other species. The recombinant BmVps4 shows ATPase activity, which can be stimulated by Mg2+ and inhibited by dominant mutations. Together, our data suggest BmVps4 is the genuine silkworm homologue of Vps4. To our knowledge, this is the first-time characterization of any silkworm MVB proteins. This study will facilitate further investigation of silkworm MVB pathway and its possible roles in the infection and budding of B. mori nuclear polyhedrosis virus (BmNPV), which is one of the most common and severe pathogens for silkworms. The cloned BmVps4 sequence is deposited in GenBank (Accession number GQ995504).