The master regulator of the cellular stress response (HSF1) is critical for orthopoxvirus infection.
The master regulator of the cellular stress response (HSF1) is critical for orthopoxvirus infection.
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DOI：
10.1371/journal.ppat.1003904
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发表时间：
2014-02
期刊：
影响因子：
6.7
通讯作者：
Connor J
中科院分区：
文献类型：
--
作者：
Filone CM;Caballero IS;Dower K;Mendillo ML;Cowley GS;Santagata S;Rozelle DK;Yen J;Rubins KH;Hacohen N;Root DE;Hensley LE;Connor J
正痘病毒科包含多种人类病原体，包括猴痘、天花和牛痘。这些病毒被认为比其他DNA病毒更少依赖宿主功能，因为它们具有大的基因组并在细胞质中复制，但缺乏对正痘病毒所需宿主因子的详细了解。为了解决这个问题，我们进行了一项无偏见的全基因组合并RNAi筛选，靶向超过17，000个人类基因，以确定支持正痘病毒感染的宿主因素。我们使用二级和三级检测来验证我们的筛选结果。最强的打击之一是热休克因子1（HSF 1），它是细胞保护热休克反应的古老主要调节因子。在研究HSF 1在牛痘感染过程中的行为时，我们发现HSF 1被磷酸化，易位到细胞核，并增加HSF 1靶基因的转录。HSF1的激活支持病毒复制，因为RNAi敲低和HSF1小分子抑制阻止正痘病毒感染。与其作为转录激活因子的作用一致，抑制几个HSF 1靶点也阻断了牛痘病毒的复制。这些数据表明，正痘病毒为了自身的利益而选择宿主转录反应，从而有效地扩展其功能基因组以包括驻留在宿主DNA内的基因。对HSF1及其分子伴侣网络的依赖性为抗病毒药物的开发提供了多种机会。正痘病毒带来许多复制所需的因子，并通过阻止宿主蛋白质的表达来损害宿主细胞。虽然正痘病毒比某些病毒对宿主的依赖性更小，但感染仍然需要宿主因素。在这里，我们报告的结果，从两个基因组规模的方法，确定宿主蛋白使用的正痘病毒在感染过程中。这些方法表明，热休克反应的主调节因子热休克因子1（HSF 1）是正痘病毒复制的关键宿主因子。HSF1调节基因是在病毒感染后表达保持或增加的仅有的宿主基因中的一些。我们的研究表明，正痘病毒进入细胞并激活宿主转录途径作为其自身复制过程的一部分。然后，这些蛋白质在感染期间被病毒利用并包装到病毒体中，基本上扩展了病毒基因组以包括从宿主核DNA中挑选的基因。这得到了非正痘病毒属的病毒基因组中存在热休克蛋白的支持。我们进一步表明，HSF1和HSF1转录基因的小分子抑制剂是正痘病毒复制的有效抑制剂，这表明了抗病毒开发的新途径。
The genus Orthopoxviridae contains a diverse group of human pathogens including monkeypox, smallpox and vaccinia. These viruses are presumed to be less dependent on host functions than other DNA viruses because they have large genomes and replicate in the cytoplasm, but a detailed understanding of the host factors required by orthopoxviruses is lacking. To address this topic, we performed an unbiased, genome-wide pooled RNAi screen targeting over 17,000 human genes to identify the host factors that support orthopoxvirus infection. We used secondary and tertiary assays to validate our screen results. One of the strongest hits was heat shock factor 1 (HSF1), the ancient master regulator of the cytoprotective heat-shock response. In investigating the behavior of HSF1 during vaccinia infection, we found that HSF1 was phosphorylated, translocated to the nucleus, and increased transcription of HSF1 target genes. Activation of HSF1 was supportive for virus replication, as RNAi knockdown and HSF1 small molecule inhibition prevented orthopoxvirus infection. Consistent with its role as a transcriptional activator, inhibition of several HSF1 targets also blocked vaccinia virus replication. These data show that orthopoxviruses co-opt host transcriptional responses for their own benefit, thereby effectively extending their functional genome to include genes residing within the host DNA. The dependence on HSF1 and its chaperone network offers multiple opportunities for antiviral drug development. Orthopoxviruses bring in many of the factors they need for replication and impair the host cell by preventing the expression of host proteins. Although orthopoxviruses are less reliant on the host than some viruses, host factors are still required for infection. Here, we report results from two genome-scale approaches that identify host proteins used by orthopoxviruses during infection. These approaches showed that the master regulator of the heat shock response, heat shock factor 1 (HSF1), is a critical host factor for orthopoxvirus replication. HSF1-regulated genes are some of the only host genes with expression maintained or increased following virus infection. Our studies show that orthopoxviruses enter the cell and activate a host transcription pathway as part of its own replication process. These proteins are then utilized by the virus during infection and packaged into the virion, essentially extending the viral genome to include genes co-opted from the host nuclear DNA. This is supported by the existence of heat shock proteins in the viral genome of non-orthopoxvirus genera. We further show that small-molecule inhibitors of HSF1 and HSF1-transcribed genes are effective inhibitors of orthopoxvirus replication, suggesting a new avenue for antiviral development.
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影响因子：
7
作者：
Harrow J;Frankish A;Gonzalez JM;Tapanari E;Diekhans M;Kokocinski F;Aken BL;Barrell D;Zadissa A;Searle S;Barnes I;Bignell A;Boychenko V;Hunt T;Kay M;Mukherjee G;Rajan J;Despacio-Reyes G;Saunders G;Steward C;Harte R;Lin M;Howald C;Tanzer A;Derrien T;Chrast J;Walters N;Balasubramanian S;Pei B;Tress M;Rodriguez JM;Ezkurdia I;van Baren J;Brent M;Haussler D;Kellis M;Valencia A;Reymond A;Gerstein M;Guigó R;Hubbard TJ
通讯作者：
Hubbard TJ
影响因子：
4.1
作者：
Andrade, AA;Silva, PNG;Bonjardim, CA
通讯作者：
Bonjardim, CA
影响因子：
64.5
作者：
Dai, Chengkai;Whitesell, Luke;Lindquist, Susan
通讯作者：
Lindquist, Susan
影响因子：
5.4
作者：
Guerra, S;López-Fernández, LA;Esteban, M
通讯作者：
Esteban, M
影响因子：
4.8
作者：
Johnston, Sara C.;Lin, Kenny L.;Hensley, Lisa E.
通讯作者：
Hensley, Lisa E.