BmNPV多角体蛋白基因编码区结合蛋白的鉴定

Identification of Binding Proteins of BmNPV polh CDS

家蚕核型多角体病毒(Bombyx mori nucleopolyhedrovirus,BmNPV)是一种具有较强传染性的病毒,在感染的极晚期超高水平表达一种约29 kD的碱溶性多角体蛋白。但该蛋白为病毒复制的非必需蛋白,因此,利用该基因可以被删除或被外源基因替换而不影响病毒的复制的原理构建重组病毒,可以实现外源基因的高水平表达。然而,目前虽已经有较多文献研究多角体蛋白高水平表达的分子机制,但迄今对于多角体蛋白基因编码区的结合蛋白尚不明确。本研究通过DNA pull-down联合质谱技术对结合在多角体蛋白编码区的蛋白质进行鉴定,共鉴定到78个宿主来源的蛋白和49个病毒自身编码的蛋白。对这些蛋白进行GO注释,结果表明这些蛋白除了具有共同的核酸结合功能外,还有其他不同的生物学功能,共同在多角体蛋白的超高水平表达过程中发挥作用。进一步从上述蛋白中筛选出了最值得深入研究的9个宿主蛋白和10个病毒蛋白。对这些蛋白进行深入研究,将为深入揭示多角体蛋白超高水平表达的分子机制提供新的线索。

Bombyx mori nucleopolyhedrosis virus is a virulent and infectious virus,which highly expresses polyhedrin,an alkali-soluble protein of about 29 kD in the very late phase.At the same time,this protein is not necessary for viral replication.Therefore,this gene can be deleted or replaced without affecting viral replication,and the promoter of this gene has been used to construct recombinant viruses to achieve hyperexpression of foreign genes.However,although a large number of studies have been carried out to explore the expression mechanism of polyhedrin,the binding proteins of the coding region of polh have not been identified.In this research,a total of 78 host proteins and 49 viral proteins bound to polh CDS were screened out by DNA pull-down combined with mass spectrometry.Apart from that,through GO annotation,it was found that in addition to the common nucleic acid binding function,the host proteins and the viral proteins had their different biological functions,and jointly played a role in the hyperexpression of polyhedrin.In order to narrow the scope of important regulatory proteins,we screened 9 host proteins and 10 viral proteins.Insights into the molecular functions of these proteins will shed new light on the molecular mechanism of hyperexpression of BmNPV polyhedrin.