Dong H-F, Wigmore K, Carrington M N, Dean M, Turpin J A, Howard O M Z
Basic Research Program, SAIC Frederick, MD 21702, USA.
Genes Immun. 2005 Oct;6(7):609-19. doi: 10.1038/sj.gene.6364247.
CCR5 is one of the primary coreceptors for Env-mediated fusion between cells and human immunodeficiency virus type 1 (HIV-1). Analyses of CCR5 variants in cohorts of HIV-1 high-risk individuals led to the identification of multiple single amino-acid substitutions, which may have functional consequences. This study focused on eight naturally occurring allelic variants located between amino-acid residues 60 and 334 of CCR5. All studied allelic variants were highly expressed on the cell surface of HEK-293 cells and permissive for HIV-1 infection. Variant G301V showed 3.5-fold increase in 50% effective concentration (EC(50)) for CCL4 (MIP 1beta) in a competitive binding assay. There was also a significant reduction in CCL5 (RANTES) EC(50) for the R223Q, A335V and Y339F variants. The most unexpected functional abnormality was exhibited by the R60S variant that exhibited a loss of ligand-induced desensitization in chemotaxis assays, but showed normal CCL4 and CCL5 binding avidity. This mutation is located in the first intracellular loop, a domain that has not previously been shown to be involved in receptor desensitization. In conclusion, our results support earlier studies showing that these naturally occurring point mutations do not limit HIV-1 infection, and indicated that single amino-acid changes can have unexpected functional consequences.
CCR5是Env介导的细胞与1型人类免疫缺陷病毒(HIV-1)之间融合的主要共受体之一。对HIV-1高危个体队列中的CCR5变体进行分析后,鉴定出多个单氨基酸取代,这些取代可能具有功能影响。本研究聚焦于位于CCR5氨基酸残基60至334之间的8个天然存在的等位基因变体。所有研究的等位基因变体均在HEK-293细胞表面高度表达,并允许HIV-1感染。在竞争性结合试验中,变体G301V对CCL4(MIP 1β)的50%有效浓度(EC50)增加了3.5倍。R223Q、A335V和Y339F变体的CCL5(RANTES)EC50也显著降低。最出乎意料的功能异常由R60S变体表现出来,该变体在趋化试验中表现出配体诱导的脱敏丧失,但CCL4和CCL5结合亲和力正常。此突变位于第一个细胞内环,该结构域以前未被证明参与受体脱敏。总之,我们的结果支持早期研究,表明这些天然存在的点突变不会限制HIV-1感染,并表明单氨基酸变化可能产生意想不到的功能后果。