The Broad-Spectrum Antiviral Protein ZAP Restricts Human Retrotransposition.
The Broad-Spectrum Antiviral Protein ZAP Restricts Human Retrotransposition.
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DOI：
10.1371/journal.pgen.1005252
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发表时间：
2015-05
期刊：
影响因子：
4.5
通讯作者：
Kazazian HH Jr
中科院分区：
文献类型：
--
作者：
Goodier JL;Pereira GC;Cheung LE;Rose RJ;Kazazian HH Jr
固有免疫描述了一组最近发现但知之甚少的细胞机制，专门针对病毒病原体。他们的发现在很大程度上源于对HIV和SIV的深入研究，这些研究揭示了在逆转录病毒生命周期的各个阶段起作用的限制因子。最近的研究表明，一些因素限制逆转录病毒和逆转录转座子，但令人惊讶的是，在方式上可能有所不同。我们筛选了已知的干扰素刺激的抗病毒蛋白，以前没有测试过它们对细胞培养逆转录转座的影响。包括BST 2、ISG 20、MAVS、MX2和ZAP在内的几个因子显示出强烈的L1抑制。我们专注于ZAP（PARP 13/ZC 3 HAV 1），一种锌指蛋白，靶向几个家族的病毒，包括逆转录病毒科，Tiloviridae和披膜病毒科，并表明ZAP表达也通过L1 RNA和核糖核蛋白颗粒完整性的丧失强烈限制细胞培养中的逆转录转座。ZAP与L1核糖核蛋白颗粒的关联得到免疫共沉淀和与ORF 1 p在细胞质应激颗粒中的共定位的支持。我们还使用质谱法来确定ZAP相互作用组的蛋白质组分，并鉴定了许多与ZAP直接相互作用和共定位的蛋白质，包括MOV 10，这是一种先前显示可抑制反转录转座子的RNA解旋酶。伴侣蛋白复合物、RNA降解蛋白、解旋酶、翻译后修饰剂和染色质修饰复合物的组分的检测表明ZAP抗逆转录酶活性的机制在细胞质中并且可能也在细胞核中起作用。ZAP核糖核蛋白颗粒与许多干扰素刺激的基因产物的关联表明它可能是干扰素应答中的关键参与者。反转录转座子是移动的DNA元件，其通过使用RNA中间体的“复制和粘贴”机制复制自身。它们是插入诱变剂，对基因组进化产生深远影响，促进DNA缺失、插入和重排，并改变基因表达。LINE-1逆转录转座子占据了人类DNA的17%，尽管据信在任何个体中只有约100个仍然具有逆转录转座子的能力。细胞已经进化出限制逆转录转座的防御，在某些情况下涉及干扰素刺激基因（ISG），这些基因是保护细胞免受病毒感染的先天免疫系统的一部分。我们筛选了一组ISG，并在细胞培养试验中发现了几种强烈限制反转录转座的ISG。我们的研究增加了对ZAP的理解，ZAP是一种重要的限制因子，可以对抗正链和负链RNA以及一些DNA病毒，也可以与人类逆转录转座子相互作用以防止基因组突变。显微镜和免疫沉淀显示ZAP蛋白与L1核糖核蛋白颗粒以及MOV 10（一种也抑制反转录转座子的RNA解旋酶）密切相关。ZAP蛋白相互作用组的详细检查揭示了许多其他ISG直接结合ZAP，并为探索ZAP介导的抗逆转录酶活性的机制提出了新的方向。
Intrinsic immunity describes the set of recently discovered but poorly understood cellular mechanisms that specifically target viral pathogens. Their discovery derives in large part from intensive studies of HIV and SIV that revealed restriction factors acting at various stages of the retroviral life cycle. Recent studies indicate that some factors restrict both retroviruses and retrotransposons but surprisingly in ways that may differ. We screened known interferon-stimulated antiviral proteins previously untested for their effects on cell culture retrotransposition. Several factors, including BST2, ISG20, MAVS, MX2, and ZAP, showed strong L1 inhibition. We focused on ZAP (PARP13/ZC3HAV1), a zinc-finger protein that targets viruses of several families, including Retroviridae, Tiloviridae, and Togaviridae, and show that ZAP expression also strongly restricts retrotransposition in cell culture through loss of L1 RNA and ribonucleoprotein particle integrity. Association of ZAP with the L1 ribonucleoprotein particle is supported by co-immunoprecipitation and co-localization with ORF1p in cytoplasmic stress granules. We also used mass spectrometry to determine the protein components of the ZAP interactome, and identified many proteins that directly interact and colocalize with ZAP, including MOV10, an RNA helicase previously shown to suppress retrotransposons. The detection of a chaperonin complex, RNA degradation proteins, helicases, post-translational modifiers, and components of chromatin modifying complexes suggest mechanisms of ZAP anti-retroelement activity that function in the cytoplasm and perhaps also in the nucleus. The association of the ZAP ribonucleoprotein particle with many interferon-stimulated gene products indicates it may be a key player in the interferon response. Retrotransposons are mobile DNA elements that duplicate themselves by a "copy and paste" mechanism using an RNA intermediate. They are insertional mutagens that have had profound effects on genome evolution, fostering DNA deletions, insertions and rearrangements, and altering gene expression. LINE-1 retrotransposons occupy 17% of human DNA, although it is believed that only about 100 remain competent for retrotransposition in any individual. The cell has evolved defenses restricting retrotransposition, involving in some cases interferon-stimulated genes (ISGs) that are part of the innate immune system that protects the cell from viral infections. We screened a panel of ISGs and found several to strongly limit retrotransposition in a cell culture assay. Our investigations increase understanding of how ZAP, an important restriction factor against positive- and negative-strand RNA and some DNA viruses, also interacts with human retrotransposons to prevent genome mutation. Microscopy and immunoprecipitation show a close association of ZAP protein with the L1 ribonucleoprotein particle, as well as MOV10, an RNA helicase that also inhibits retrotransposons. A detailed examination of the ZAP protein interactome reveals many other ISGs that directly bind ZAP, and suggests new directions for exploring the mechanisms of ZAP-mediated anti-retroelement activity.
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影响因子：
5.2
作者：
Arias JF;Koyama T;Kinomoto M;Tokunaga K
通讯作者：
Tokunaga K
影响因子：
3.3
作者：
Arjan-Odedra S;Swanson CM;Sherer NM;Wolinsky SM;Malim MH
通讯作者：
Malim MH
DOI：
10.1073/pnas.0831042100
发表时间：
2003-04-29
影响因子：
11.1
作者：
Brouha, B;Schustak, J;Kazazian, HH
通讯作者：
Kazazian, HH
影响因子：
30.8
作者：
Dewannieux, M;Esnault, C;Heidmann, T
通讯作者：
Heidmann, T
DOI：
10.1073/pnas.1302564110
发表时间：
2013-07-02
影响因子：
11.1
作者：
Charron, Guillaume;Li, Melody M. H.;Hang, Howard C.
通讯作者：
Hang, Howard C.