German Primate Center - Leibniz Institute for Primate Research, Göttingen, Germany.
Universitätsklinikum Erlangen, Institute for Clinical and Molecular Virology, Erlangen, Germany.
PLoS Pathog. 2021 Mar 3;17(3):e1008979. doi: 10.1371/journal.ppat.1008979. eCollection 2021 Mar.
The rhesus monkey rhadinovirus (RRV), a γ2-herpesvirus of rhesus macaques, shares many biological features with the human pathogenic Kaposi's sarcoma-associated herpesvirus (KSHV). Both viruses, as well as the more distantly related Epstein-Barr virus, engage cellular receptors from the Eph family of receptor tyrosine kinases (Ephs). However, the importance of the Eph interaction for RRV entry varies between cell types suggesting the existence of Eph-independent entry pathways. We therefore aimed to identify additional cellular receptors for RRV by affinity enrichment and mass spectrometry. We identified an additional receptor family, the Plexin domain containing proteins 1 and 2 (Plxdc1/2) that bind the RRV gH/gL glycoprotein complex. Preincubation of RRV with soluble Plxdc2 decoy receptor reduced infection by ~60%, while overexpression of Plxdc1 and 2 dramatically enhanced RRV susceptibility and cell-cell fusion of otherwise marginally permissive Raji cells. While the Plxdc2 interaction is conserved between two RRV strains, 26-95 and 17577, Plxdc1 specifically interacts with RRV 26-95 gH. The Plxdc interaction is mediated by a short motif at the N-terminus of RRV gH that is partially conserved between isolate 26-95 and isolate 17577, but absent in KSHV gH. Mutation of this motif abrogated the interaction with Plxdc1/2 and reduced RRV infection in a cell type-specific manner. Taken together, our findings characterize Plxdc1/2 as novel interaction partners and entry receptors for RRV and support the concept of the N-terminal domain of the gammaherpesviral gH/gL complex as a multifunctional receptor-binding domain. Further, Plxdc1/2 usage defines an important biological difference between KSHV and RRV.
恒河猴疱疹病毒(RRV)是恒河猴的一种γ2疱疹病毒,与人类致病的卡波西肉瘤相关疱疹病毒(KSHV)有许多生物学特征。这两种病毒以及更为遥远相关的 EBV,都与细胞受体酪氨酸激酶(Ephs)家族的 Eph 受体结合。然而,Eph 相互作用对 RRV 进入的重要性因细胞类型而异,这表明存在 Eph 非依赖性进入途径。因此,我们旨在通过亲和富集和质谱法鉴定 RRV 的其他细胞受体。我们鉴定了另一个受体家族,即包含 Plexin 结构域的蛋白 1 和 2(Plxdc1/2),它们与 RRV gH/gL 糖蛋白复合物结合。RRV 与可溶性 Plxdc2 诱饵受体预孵育可将感染减少约 60%,而 Plxdc1 和 2 的过表达则显著增强了原本允许度较低的 Raji 细胞的 RRV 易感性和细胞-细胞融合。虽然 Plxdc2 相互作用在两种 RRV 株(26-95 和 17577)之间是保守的,但 Plxdc1 仅与 RRV 26-95 gH 相互作用。Plxdc 相互作用由 RRV gH 的 N 端短基序介导,该基序在 26-95 分离株和 17577 分离株之间部分保守,但在 KSHV gH 中缺失。该基序的突变消除了与 Plxdc1/2 的相互作用,并以细胞类型特异性的方式降低了 RRV 感染。总之,我们的研究结果将 Plxdc1/2 鉴定为 RRV 的新型相互作用伙伴和进入受体,并支持γ-疱疹病毒 gH/gL 复合物的 N 端结构域作为多功能受体结合域的概念。此外,Plxdc1/2 的使用定义了 KSHV 和 RRV 之间的一个重要生物学差异。