Rai Tia, Marble Deborah, Rihani Kayla, Rong Lijun
Department of Microbiology and Immunology, College of Medicine, University of Illinois at Chicago, Chicago, Illinois 60612, USA.
J Virol. 2004 Jan;78(2):683-91. doi: 10.1128/jvi.78.2.683-691.2004.
Rong et al. have demonstrated previously that with a few substitutions, the fourth repeat of human low-density lipoprotein (hLDL-A4) receptor can functionally replace the LDL-A module of Tva, the cellular receptor for subgroup A avian sarcoma and leukosis virus (ASLV-A), in viral entry (L. Rong, K. Gendron, and P. Bates, Proc. Natl. Acad. Sci. USA 95:8467-8472, 1998). Here we have shown that swapping the amino terminus of hLDL repeat 5 (hLDL-A5) with that of Tva, in addition to the corresponding substitutions made in human LDL-A4, was required to convert hLDL-A5 into an efficient ASLV-A receptor. These results substantiated our previous findings regarding the role of the specific residues in the viral interaction domain of Tva and demonstrated the critical role of the amino terminus of the Tva LDL-A module in ASLV-A infection. Furthermore, we have shown that the residues between cysteines 2 and 3 of the Tva LDL-A module in a Tva/LDL-A5 chimeric protein can be functionally replaced by the corresponding region of another LDL-A module, human LDL receptor-related protein repeat 22 (LDL-A22), to mediate efficient ASLV-A entry. Since the only conserved feature between the C2-C3 region of LDL-A22 and the Tva LDL-A module is that both contain nine amino acids of which none are conserved, we conclude that the spacing between C2 and C3 of the LDL-A module of Tva is an important determinant for ASLV-A entry. Thus, the present study provides strong evidence to support our hypothesis that one role of the N terminus of the LDL-A module of Tva is to allow proper folding and conformation of the protein for optimal interaction with the viral glycoprotein EnvA in ASLV-A entry.
荣等人先前已证明,通过一些替换,人类低密度脂蛋白(hLDL-A4)受体的第四个重复序列在病毒进入过程中能够功能性地替代Tva的LDL-A模块,Tva是A亚群禽肉瘤和白血病病毒(ASLV-A)的细胞受体(L.荣、K.根德龙和P.贝茨,《美国国家科学院院刊》95:8467 - 8472,1998)。在此我们表明,除了在人类LDL-A4中进行相应替换外,将hLDL重复序列5(hLDL-A5)的氨基末端与Tva的氨基末端进行交换,是将hLDL-A5转化为高效ASLV-A受体所必需的。这些结果证实了我们先前关于Tva病毒相互作用结构域中特定残基作用的发现,并证明了Tva LDL-A模块氨基末端在ASLV-A感染中的关键作用。此外,我们还表明,在Tva/LDL-A5嵌合蛋白中,Tva LDL-A模块第2和第3个半胱氨酸之间的残基可以被另一个LDL-A模块,即人类低密度脂蛋白受体相关蛋白重复序列22(LDL-A22)的相应区域功能性替代,以介导高效的ASLV-A进入。由于LDL-A22的C2 - C3区域与Tva LDL-A模块之间唯一保守的特征是两者都包含九个氨基酸且无一个保守,我们得出结论,Tva LDL-A模块C2和C3之间的间距是ASLV-A进入的重要决定因素。因此,本研究提供了有力证据支持我们的假设,即Tva LDL-A模块氨基末端的一个作用是使蛋白质正确折叠并形成构象,以便在ASLV-A进入过程中与病毒糖蛋白EnvA进行最佳相互作用。