The Zinc Finger Antiviral Protein ZAP Restricts Human Cytomegalovirus and Selectively Binds and Destabilizes Viral UL4/UL5 Transcripts.
The Zinc Finger Antiviral Protein ZAP Restricts Human Cytomegalovirus and Selectively Binds and Destabilizes Viral UL4/UL5 Transcripts.
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DOI：
10.1128/mbio.02683-20
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发表时间：
2021-05-04
期刊：
影响因子：
6.4
通讯作者：
Brinkmann MM
中科院分区：
文献类型：
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作者：
Gonzalez-Perez AC;Stempel M;Wyler E;Urban C;Piras A;Hennig T;Ganskih S;Wei Y;Heim A;Landthaler M;Pichlmair A;Dölken L;Munschauer M;Erhard F;Brinkmann MM
干扰素刺激的基因产物（ISGs）在早期感染控制中起着至关重要的作用。ISG锌指CCCH型抗病毒蛋白1（ZAP/ZC 3 HAV 1）通过与富含CG的RNA序列结合来拮抗几种RNA病毒，而其对DNA病毒的作用尚不清楚。在这里，我们破译了ZAP在人巨细胞病毒（HCMV）感染背景下的作用，人巨细胞病毒（HCMV）是一种与免疫抑制个体和新生儿的高发病率相关的β-疱疹病毒。我们表明，ZAP的两个主要亚型，ZAP-S和ZAP-L的表达，诱导HCMV感染期间，都有负面影响HCMV复制。转录组和蛋白质组分析表明，ZAP的表达导致病毒mRNA和蛋白质水平降低，并减缓HCMV感染的进展。代谢RNA标记结合高通量测序（SLAM-seq）显示，感染后期的大多数基因表达变化是由HCMV的一般减毒引起的。此外，在感染的早期阶段，ZAP通过使病毒mRNA的不同子集不稳定来限制HCMV，特别是来自先前未表征的UL 4-UL 6 HCMV基因座的那些。通过增强的交联免疫沉淀和测序分析（eCLIP-seq），我们鉴定了从该HCMV位点表达的转录物作为ZAP的直接靶点。此外，我们的数据表明，ZAP优先识别不仅CG，而且其他胞嘧啶丰富的序列，从而扩大其目标特异性。总之，本报告首次揭示了ZAP在HCMV感染过程中的直接靶点，这强烈表明来自UL 4-UL 6位点的转录物可能在HCMV复制中起重要作用。
Interferon-stimulated gene products (ISGs) play a crucial role in early infection control. The ISG zinc finger CCCH-type antiviral protein 1 (ZAP/ZC3HAV1) antagonizes several RNA viruses by binding to CG-rich RNA sequences, whereas its effect on DNA viruses is less well understood. Here, we decipher the role of ZAP in the context of human cytomegalovirus (HCMV) infection, a β-herpesvirus that is associated with high morbidity in immunosuppressed individuals and newborns. We show that expression of the two major isoforms of ZAP, ZAP-S and ZAP-L, is induced during HCMV infection and that both negatively affect HCMV replication. Transcriptome and proteome analyses demonstrated that the expression of ZAP results in reduced viral mRNA and protein levels and decelerates the progression of HCMV infection. Metabolic RNA labeling combined with high-throughput sequencing (SLAM-seq) revealed that most of the gene expression changes late in infection result from the general attenuation of HCMV. Furthermore, at early stages of infection, ZAP restricts HCMV by destabilizing a distinct subset of viral mRNAs, particularly those from the previously uncharacterized UL4-UL6 HCMV gene locus. Through enhanced cross-linking immunoprecipitation and sequencing analysis (eCLIP-seq), we identified the transcripts expressed from this HCMV locus as the direct targets of ZAP. Moreover, our data show that ZAP preferentially recognizes not only CG, but also other cytosine-rich sequences, thereby expanding its target specificity. In summary, this report is the first to reveal direct targets of ZAP during HCMV infection, which strongly indicates that transcripts from the UL4-UL6 locus may play an important role for HCMV replication.