Katsuma Susumu, Shimada Toru, Kobayashi Masahiko
Department of Agricultural and Environmental Biology, Graduate School of Agricultural and Life Sciences, The University of Tokyo, Yayoi 1-1-1, Bunkyo-ku, Tokyo 113-8657, Japan.
Virus Genes. 2004 Oct;29(2):211-7. doi: 10.1023/B:VIRU.0000036381.11779.dd.
We characterized a gene of the baculovirus Bombyx mori nucleopolyhedrovirus (BmNPV) homologous to the mammalian fibroblast growth factor (FGF) family. We termed it vfgf, and examined its transcription and the properties of the gene product (vFGF). RT-PCR analysis showed that vfgf is one of the baculovirus early genes, although there are no consensus sequences of the baculovirus early gene promoters. 5'-RACE analysis revealed that its transcription started at 10 nucleotides upstream of the translation start codon. vFGF has a hydrophobic amino terminus (approximately 16 amino acids), which is a typical signal sequence. As expected, vFGF was efficiently secreted from BmNPV-infected BmN cells. Because possible glycosylation sites are found at positions 44 (Asn) and 171 (Asn), we examined whether BmNPV vFGF is glycosylated or not. Cleavage of recombinant vFGF with PNGase F revealed that BmNPV vFGF was glycosylated. We also found that secretion of vFGF is completely blocked by the treatment of Tunicamycin, which blocks N-linked glycosylation. This is the first report to characterize a virus-encoded FGF.
我们鉴定了一种与哺乳动物成纤维细胞生长因子(FGF)家族同源的家蚕核型多角体病毒(BmNPV)基因。我们将其命名为vfgf,并检测了其转录情况以及基因产物(vFGF)的特性。逆转录聚合酶链反应(RT-PCR)分析表明,vfgf是杆状病毒早期基因之一,尽管杆状病毒早期基因启动子没有共有序列。5'-末端快速扩增(5'-RACE)分析显示,其转录起始于翻译起始密码子上游10个核苷酸处。vFGF具有一个疏水的氨基末端(约16个氨基酸),这是一个典型的信号序列。正如预期的那样,vFGF从受BmNPV感染的BmN细胞中有效分泌。由于在第44位(天冬酰胺)和第171位(天冬酰胺)发现了可能的糖基化位点,我们检测了BmNPV vFGF是否被糖基化。用肽-N-糖苷酶F(PNGase F)切割重组vFGF表明BmNPV vFGF被糖基化。我们还发现,衣霉素处理可完全阻断vFGF的分泌,衣霉素可阻断N-连接糖基化。这是首篇对病毒编码的FGF进行鉴定的报道。