Section on Molecular Virology, Laboratory of Cellular and Molecular Regulation, National Institute of Mental Health, National Institutes of Health, Bethesda, MD 20892, USA.
J Virol. 2011 Apr;85(7):3498-506. doi: 10.1128/JVI.02586-10. Epub 2011 Jan 26.
BHK cells remain resistant to xenotropic murine retrovirus-related virus (XMRV) or gibbon ape leukemia virus (GALV) infection, even when their respective receptors, Xpr1 or PiT1, are expressed. We set out to determine the stage at which viral infection is blocked and whether this block is mediated by a dominant-negative factor or the absence of a requisite ancillary factor. BHK cells bind neither XMRV nor GALV envelope proteins. BHK cells expressing the appropriate receptors bind XMRV or GALV envelope proteins. BHK cells can be infected by NZB-XMV(New Zealand Black mouse xenotropic murine virus)-enveloped vectors, expressing an envelope derived from a xenotropic retrovirus that, like XMRV, employs Xpr1 as a receptor, and also by vectors bearing the envelope of 10A1 murine leukemia virus (MLV), a murine retrovirus that can use PiT1 as a receptor. The retroviral vectors used in these analyses differ solely in their viral envelope proteins, suggesting that the block to XMRV and GALV infection is mediated at the level of envelope-receptor interactions. N-linked glycosylation of the receptors was not found to mediate resistance of receptor-expressing BHK cells to GALV or XMRV, as shown by tunicamycin treatment and mutation of the specific glycosylation site of the PiT1 receptor. Hybrid cells produced by fusing BHKXpr1 or BHKPiT1 to XMRV- or GALV-resistant cells, respectively, can mediate efficient XMRV or GALV infection. These findings indicate that BHK cells lack a factor that is required for infection by primate xenotropic viruses. This factor is not required for viruses that use the same receptors but were directly isolated from mice.
BHK 细胞仍然对异嗜性鼠白血病病毒相关病毒 (XMRV) 或长臂猿白血病病毒 (GALV) 感染具有抗性,即使表达了它们各自的受体 Xpr1 或 PiT1 也是如此。我们着手确定病毒感染被阻断的阶段,以及这种阻断是否是由显性负性因子或缺乏必需辅助因子介导的。BHK 细胞既不结合 XMRV 也不结合 GALV 包膜蛋白。表达适当受体的 BHK 细胞结合 XMRV 或 GALV 包膜蛋白。BHK 细胞可以被表达源自类似于 XMRV 的异嗜性逆转录病毒的包膜的 NZB-XMV(新西兰黑鼠异嗜性鼠白血病病毒)包膜载体感染,也可以被携带 10A1 鼠白血病病毒 (MLV) 包膜的载体感染,10A1 鼠白血病病毒是一种可以使用 PiT1 作为受体的鼠逆转录病毒。这些分析中使用的逆转录病毒载体仅在其病毒包膜蛋白上有所不同,这表明 XMRV 和 GALV 感染的阻断是在包膜-受体相互作用的水平上介导的。如用衣霉素处理和突变 PiT1 受体的特定糖基化位点所示,受体的 N 连接糖基化并未介导表达受体的 BHK 细胞对 GALV 或 XMRV 的抗性。通过分别将 BHKXpr1 或 BHKPiT1 与 XMRV 或 GALV 抗性细胞融合产生的杂交细胞可以介导有效的 XMRV 或 GALV 感染。这些发现表明 BHK 细胞缺乏感染灵长类异嗜性病毒所需的因子。该因子对于使用相同受体但直接从小鼠中分离出来的病毒不是必需的。