Macroautophagy/autophagy is an auto-digestive pro-survival pathway activated in response to stress to target cargo for lysosomal degradation. In recent years, autophagy has become prominent as an innate antiviral defense mechanism through multiple processes, such as targeting virions and viral components for elimination. These exciting findings have encouraged studies on the ability of autophagy to restrict HIV. However, the role of autophagy in HIV infection remains unclear. Whereas some reports indicate that autophagy is detrimental for HIV, others have claimed that HIV deliberately activates this pathway to increase its infectivity. Moreover, these contrasting findings seem to depend on the cell type investigated. Here, we show that autophagy poses a hurdle for HIV replication, significantly reducing virion production. However, HIV-1 uses its accessory protein Nef to counteract this restriction. Previous studies have indicated that Nef affects autophagy maturation by preventing the fusion between autophagosomes and lysosomes. Here, we uncover that Nef additionally blocks autophagy initiation by enhancing the association between BECN1 and its inhibitor BCL2, and this activity depends on the cellular E3 ligase PRKN. Remarkably, the ability of Nef to counteract the autophagy block is more frequently observed in pandemic HIV-1 and its simian precursor SIVcpz infecting chimpanzees than in HIV-2 and its precursor SIVsmm infecting sooty mangabeys. In summary, our findings demonstrate that HIV-1 is susceptible to autophagy restriction and define Nef as the primary autophagy antagonist of this antiviral process.Abbreviations: 3-MA: 3-methyladenine; ACTB: actin, beta; ATG16L1: autophagy related 16 like 1; BCL2: bcl2 apoptosis regulator; BECN1: beclin 1; cDNA: complementary DNA; EGFP: enhanced green fluorescence protein; ER: endoplasmic reticulum; Gag/p55: group-specific antigen; GFP: green fluorescence protein; GST: glutathione S transferase; HA: hemagglutinin; HIV: human immunodeficiency virus; IP: immunoprecipitation; MAP1LC3B/LC3B: microtubule associated protein 1 light chain 3 beta; Nef: negative factor; PRKN: parkin RBR E3 ubiquitin ligase; PtdIns3K: phosphatidylinositol 3 kinase; PtdIns3P: phosphatidylinositol 3 phosphate; PTM: post-translational modification; RT-qPCR: reverse transcription followed by quantitative PCR; RUBCN: rubicon autophagy regulator; SEM: standard error of the mean; SERINC3: serine incorporator 3; SERINC5: serine incorporator 5; SIV: simian immunodeficiency virus; SQSTM1/p62: sequestosome 1; TFEB: transcription factor EB; UVRAG: UV radiation resistance associated gene; VSV: vesicular stomatitis virus; ZFYVE1/DFCP1: zinc finger FYVE-type containing 1.

中文翻译：

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HIV-1 Nef 通过以 PRKN 依赖性方式增强 BECN1 与其抑制剂 BCL2 之间的关联来抵消自噬限制。

巨自噬/自噬是一种自动消化的促生存途径，响应于对目标货物进行溶酶体降解的应激而激活。近年来，自噬作为一种先天抗病毒防御机制，通过多种过程（例如针对病毒粒子和病毒成分进行消除）而变得突出。这些令人兴奋的发现鼓励了对自噬限制艾滋病毒能力的研究。然而，自噬在 HIV 感染中的作用仍不清楚。虽然一些报告表明自噬对 HIV 有害，但其他报告则声称 HIV 故意激活该途径以增加其传染性。此外，这些对比的发现似乎取决于所研究的细胞类型。在这里，我们发现自噬对 HIV 复制构成了障碍，显着减少了病毒颗粒的产生。然而，HIV-1 使用其辅助蛋白 Nef 来抵消这种限制。先前的研究表明，Nef 通过阻止自噬体和溶酶体之间的融合来影响自噬成熟。在这里，我们发现 Nef 通过增强 BECN1 与其抑制剂 BCL2 之间的关联来额外阻止自噬启动，并且这种活性取决于细胞 E3 连接酶 PRKN。值得注意的是，Nef 抵抗自噬阻断的能力在大流行的 HIV-1 及其感染黑猩猩的猿猴前体 SIVcpz 中比在感染 HIV-2 及其前体 SIVsmm 的乌白眉猴中更常见。总之，我们的研究结果表明，HIV-1 容易受到自噬限制，并将 Nef 定义为该抗病毒过程的主要自噬拮抗剂。缩写：3-MA：3-甲基腺嘌呤；3-MA：3-甲基腺嘌呤；ACTB：肌动蛋白，β；ATG16L1：自噬相关16样1；BCL2：bcl2凋亡调节因子；BECN1: 贝克林 1; cDNA：互补DNA；EGFP：增强型绿色荧光蛋白；ER：内质网；Gag/p55：群体特异性抗原；GFP：绿色荧光蛋白；GST：谷胱甘肽S转移酶；HA：血凝素；HIV：人类免疫缺陷病毒；IP：免疫沉淀；MAP1LC3B/LC3B：微管相关蛋白1轻链3β；Nef：负面因素；PRKN：parkin RBR E3 泛素连接酶；PtdIns3K：磷脂酰肌醇 3 激酶；PtdIns3P：磷脂酰肌醇3磷酸酯；PTM：翻译后修饰；RT-qPCR：逆转录后进行定量 PCR；RUBCN：rubicon自噬调节因子；SEM：平均值的标准误差；SERINC3：丝氨酸掺入子3；SERINC5：丝氨酸掺入子5；SIV：猿猴免疫缺陷病毒；SQSTM1/p62: 隔离体 1; TFEB：转录因子EB；UVRAG：抗紫外线辐射相关基因；VSV：水泡性口炎病毒；ZFYVE1/DFCP1：锌指FYVE型，含1。