Molecular cloning, expression and characterization of a novel vacuolar protein sorting 4 gene in silkworm, Bombyx mori.
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 Mg(2+) 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).
Institute of Life Sciences, Jiangsu University, 301 Xuefu Road, Zhenjiang, Jiangsu 212013, People's Republic of China.
Hengchuan
H
Xia
Chunxia
C
Zhang
Fan
F
Feng
Yi
Y
Yuan
Yang
Y
Zhou
Xiaoyong
X
Liu
Keming
K
Zhu
Qin
Q
Yao
Keping
K
Chen
D000251:Adenosine Triphosphatases / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics* / Q000302:isolation & purification; D000595:Amino Acid Sequence; D019943:Amino Acid Substitution; D000818:Animals; D001483:Base Sequence; D012831:Bombyx / Q000235:genetics*; D003001:Cloning, Molecular; D017124:Conserved Sequence; D056827:Endosomal Sorting Complexes Required for Transport / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics* / Q000302:isolation & purification; D004926:Escherichia coli; D015870:Gene Expression; D019476:Insect Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics* / Q000302:isolation & purification; D008958:Models, Molecular; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D010802:Phylogeny; D017433:Protein Structure, Secondary; D017434:Protein Structure, Tertiary; D017422:Sequence Analysis, DNA; D040681:Structural Homology, Protein
D056827:Endosomal Sorting Complexes Required for Transport; D019476:Insect Proteins; D000251:Adenosine Triphosphatases