Goujon Caroline, Arfi Vanessa, Pertel Thomas, Luban Jeremy, Lienard Julia, Rigal Dominique, Darlix Jean-Luc, Cimarelli Andrea
LaboRetro, Department of Human Virology, Ecole Normale Supérieure de Lyon, Lyon, France.
J Virol. 2008 Dec;82(24):12335-45. doi: 10.1128/JVI.01181-08. Epub 2008 Oct 1.
Human immunodeficiency virus type 2 (HIV-2)/simian immunodeficiency virus SIV(SM) Vpx is incorporated into virion particles and is thus present during the early steps of infection, when it has been reported to influence the nuclear import of viral DNA. We recently reported that Vpx promoted the accumulation of full-length viral DNA following the infection of human monocyte-derived dendritic cells (DCs). This positive effect was exerted following the infection of DCs with cognate viruses and with retroviruses as divergent as HIV-1, feline immunodeficiency virus, and even murine leukemia virus, leading us to suggest that Vpx counteracted an antiviral restriction present in DCs. Here, we show that Vpx is required, albeit to a different extent, for the infection of all myeloid but not of lymphoid cells, including monocytes, macrophages, and monocytoid THP-1 cells that had been induced to differentiate with phorbol esters. The intracellular localization of Vpx was highly heterogeneous and cell type dependent, since Vpx localized differently in HeLa cells and DCs. Despite these differences, no clear correlation between the functionality of Vpx and its intracellular localization could be drawn. As a first insight into its function, we determined that SIV(SM)/HIV-2 and SIV(RCM) Vpx proteins interact with the DCAF1 adaptor of the Cul4-based E3 ubiquitin ligase complex recently described to associate with HIV-1 Vpr and HIV-2 Vpx. However, the functionality of Vpx proteins in the infection of DCs did not strictly correlate with DCAF1 binding, and knockdown experiments failed to reveal a functional role for this association in differentiated THP-1 cells. Lastly, when transferred in the context of a replication-competent viral clone, Vpx was required for replication in DCs.
2型人类免疫缺陷病毒(HIV-2)/猴免疫缺陷病毒SIV(SM)的Vpx蛋白被整合到病毒粒子中,因此在感染的早期阶段就存在,据报道此时它会影响病毒DNA的核输入。我们最近报道,Vpx在人单核细胞衍生的树突状细胞(DC)感染后促进全长病毒DNA的积累。在用同源病毒以及与HIV-1、猫免疫缺陷病毒甚至鼠白血病病毒一样不同源的逆转录病毒感染DC后,都会产生这种积极作用,这使我们推测Vpx抵消了DC中存在的一种抗病毒限制。在此,我们表明,对于所有髓系细胞而非淋巴系细胞的感染,Vpx都是必需的,包括单核细胞、巨噬细胞以及用佛波酯诱导分化的单核细胞样THP-1细胞。Vpx的细胞内定位高度异质且依赖细胞类型,因为Vpx在HeLa细胞和DC中的定位不同。尽管存在这些差异,但Vpx的功能与其细胞内定位之间并未发现明显的相关性。作为对其功能的初步了解,我们确定SIV(SM)/HIV-2和SIV(RCM)的Vpx蛋白与基于Cul4的E3泛素连接酶复合物的DCAF1衔接蛋白相互作用,最近有报道称该复合物与HIV-1 Vpr和HIV-2 Vpx相关。然而,Vpx蛋白在DC感染中的功能与DCAF1结合并不严格相关,并且敲低实验未能揭示这种结合在分化的THP-1细胞中的功能作用。最后,当在具有复制能力的病毒克隆背景下转移时,Vpx是在DC中复制所必需的。