Identification of the functional domains in an orphan G protein-coupled receptor, GPR1, which acts as a coreceptor for the brain-derived cell tropism of HIV-1, HIV-2, and SIV

鉴定孤儿 G 蛋白偶联受体 GPR1 的功能域，该受体充当 HIV-1、HIV-2 和 SIV 脑源性细胞趋向性的辅助受体

基本信息

批准号：
12470068
负责人：
SHIMIZU Nobuaki
金额：
$ 3.2万
依托单位：
GUNMA UNIVERSITY SCHOOL OF MEDICINE
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
2000
资助国家：
日本
起止时间：
2000 至 2001
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-12470068/
关键词：
HIV-1 CORECEPTOR GPCR CELL TROPISM FUNCTIONAL DOMAIN GPCR HIV AIDS脳症 GPR1

项目摘要

【Aim】 Pericytes located around the brain blood vessels constitute the blood-brain barrier with endothelial cells. The infection of the cells with human immunodeficiency virus type-1 (HIV-1) is thought to be a causative factor to develop acquired immune deficiency syndrome (AIDS)-related encephalitis. An orphan G protein-coupled receptor, GPR1, acts as a coreceptor for HIV-1 strains which infect the brain vessels-derived pericytes. The aim of this study was to identify the important domain of GPR1 for its coreceptor activity. 【Methods】 (1) The amino-terminal domain (NTR) and three extracellular loops (ECLs) interact with HIV-1. Therefore, the DNA fragments of GPR1 coding four extracellular domains were prepared by polymerase chain reaction (PCR) and recombined with CCR5 which acts as a coreceptor for macrophage tropic HIV-1 strains. (2) Some nucleotide changes were introduced into GPR1 gene by PCR to make the amino acid deletion or substitution mutants of the NTR. (3) The chimeras betwe … More en GPR1 and CCR5 and the GPR1 mutants of the NTR were introduced into a glioma-derived cell line NP-2/CD4 which is strictly resistant to the infection with HIV-1, HIV type-2 (HIV-2), and simian immunodeficiency virus (SIV) even though the expression of CD4 is detected. Susceptibility of the established cells to various HIV-1, HIV-2, and SIV strains were detected. 【Results】 (1) All of ELC chimeras lost its coreceptor activities. (2) The NTR chimeras acted as the coreceptor. (3) The amino acid mutant of GPR1 in which the first tyrosine in the NTR was substituted with alanine lost the coreceptor function. On the other hand, the substitutions of the second, the third, and the fourth tyrosine with alanine had no effects on the function. (4) The deletion of the amino-terminal 11 amino acids of the NTR had no effects on the coreceptor activity of GPR1. 【Conclusion】 The NTR is important for the coreceptor function of GPR1. The region containing four tyrosine residues in the NTR is important for its coreceptor functions. These results are available to know the mechanisms of the HIV-1 infection to the brain vessels pericytes and applicable to the development of anti-HIV-1 drugs. Less

[AIM]位于脑血管周围的周细胞构成内皮细胞的血脑屏障。人类免疫缺陷病毒类型1（HIV-1）的细胞感染被认为是发展获得性免疫缺陷综合征（AIDS）相关的脑炎的关键因素。孤儿G蛋白偶联的受体GPR1充当感染了脑血管衍生的周细胞的HIV-1菌株的共肽。这项研究的目的是确定GPR1的重要领域的共感受器活性。 [方法]（1）氨基末端结构域（NTR）和三个细胞外回路（ECLS）与HIV-1相互作用。因此，通过聚合酶链反应（PCR）制备编码四个细胞外域的GPR1的DNA片段，并与CCR5重新组合，CCR5充当巨噬细胞热带HIV-1菌株的共肽。（2）PCR将一些核变化引入了GPR1基因中，以使NTR的氨基酸缺失或取代突变体。（3）将…更多的EN GPR1和CCR5与NTR的GPR1突变体之间的嵌合体引入了神经胶质瘤衍生的细胞系NP-2/CD4中，该细胞系NP-2/CD4严格抗HIV-1，HIV-1，HIV类型-2（HIV-2）和象征性免疫缺陷病毒（SIV），即使检测到CD4的表达，也对感染了HIV-1，HIV-1型（HIV-2）（HIV-2）和象征性免疫缺陷病毒（SIV）。检测到已建立细胞对各种HIV-1，HIV-2和SIV菌株的敏感性。 [结果]（1）所有ELC Chimeras失去了其共核活性。（2）NTR嵌合体充当共感染者。（3）GPR1的氨基酸突变体，其中NTR中的第一个酪氨酸被丙氨酸取代，失去了共感受器功能。另一方面，第二，第三和第四酪氨酸的取代对该功能没有影响。（4）NTR的氨基末端11氨基酸的缺失对GPR1的共感受器活性没有影响。 [结论] NTR对GPR1的共肽功能很重要。 NTR中含有四个酪氨酸的区域对于其对果受体的功能很重要。这些结果可用于了解HIV-1感染对脑血管周细胞的机制，并适用于抗HIV-1药物的发展。较少的