G protein-coupled receptor kinase 2 promotes flaviviridae entry and replication.
G protein-coupled receptor kinase 2 promotes flaviviridae entry and replication.
复制标题
DOI：
10.1371/journal.pntd.0001820
复制
发表时间：
2012
影响因子：
3.8
通讯作者：
Garcia-Blanco MA
中科院分区：
文献类型：
--
作者：
Le Sommer C;Barrows NJ;Bradrick SS;Pearson JL;Garcia-Blanco MA
黄病毒会引发从脑炎到出血热等多种严重疾病。发现调控感染细胞中黄病毒命运的宿主因子，可能有助于深入了解感染的分子机制，从而促进抗黄病毒药物的研发。我们进行了全基因组规模的小干扰RNA（siRNA）筛选，以发现黄热病毒（YFV）增殖所需的人类宿主因子。采用2×2的siRNA池筛选形式以及重复筛选，我们确定了一份可信度较高的YFV宿主因子清单。为了找出黄病毒之间的共性，我们将这些候选因子与先前确定的西尼罗河病毒（WNV）和登革热病毒（DENV）的宿主因子进行了比较。这种比较凸显了G蛋白偶联受体激酶家族（GRKs）对于黄病毒感染的潜在必要性。YFV宿主候选因子GRK2（也称为ADRBK1）在siRNA介导的HuH - 7细胞敲低实验以及GRK - / - 小鼠胚胎成纤维细胞中都得到了验证。此外，我们发现GRK2是登革热病毒和丙型肝炎病毒（HCV）高效增殖所必需的，这表明GRK2的需求在整个黄病毒科中是保守的。最后，我们发现GRK2通过促进病毒进入和RNA合成，参与了黄病毒生命周期的多个不同阶段。总之，我们的研究结果确定GRK2是黄病毒感染的一种新型调控因子，并表明抑制GRK2的功能可能构成治疗黄病毒相关疾病的一种新方法。
黄病毒属包括几种新发和再发病毒，如登革热病毒和黄热病毒，它们会在人类中引发严重疾病，且目前尚无获批的治疗方法。黄病毒通过节肢动物传播给人类，它们依赖大量的脊椎动物和无脊椎动物因子在这些不同的宿主中进行复制。确定参与病毒增殖的宿主因子对于理解感染的分子机制以及开发新的治疗方法至关重要。为了确定黄热病毒增殖所需的人类宿主因子，我们完成了两次全基因组规模的siRNA筛选。在发现的候选因子中有G蛋白偶联受体激酶GRK2和GRK4。我们重点关注了蛋白质GRK2，这是一种最初因其在细胞信号转导中的作用而被发现的激酶。我们发现GRK2是黄热病毒、登革热病毒和丙型肝炎病毒有效感染所需的宿主因子，并且对于病毒进入和病毒基因组的高效复制都是必需的。GRKs被认为是可成药的靶点，可用于开发广谱抗黄病毒药物。
Flaviviruses cause a wide range of severe diseases ranging from encephalitis to hemorrhagic fever. Discovery of host factors that regulate the fate of flaviviruses in infected cells could provide insight into the molecular mechanisms of infection and therefore facilitate the development of anti-flaviviral drugs. We performed genome-scale siRNA screens to discover human host factors required for yellow fever virus (YFV) propagation. Using a 2×2 siRNA pool screening format and a duplicate of the screen, we identified a high confidence list of YFV host factors. To find commonalities between flaviviruses, these candidates were compared to host factors previously identified for West Nile virus (WNV) and dengue virus (DENV). This comparison highlighted a potential requirement for the G protein-coupled receptor kinase family, GRKs, for flaviviral infection. The YFV host candidate GRK2 (also known as ADRBK1) was validated both in siRNA-mediated knockdown HuH-7 cells and in GRK−/− mouse embryonic fibroblasts. Additionally, we showed that GRK2 was required for efficient propagation of DENV and Hepatitis C virus (HCV) indicating that GRK2 requirement is conserved throughout the Flaviviridae. Finally, we found that GRK2 participates in multiple distinct steps of the flavivirus life cycle by promoting both entry and RNA synthesis. Together, our findings identified GRK2 as a novel regulator of flavivirus infection and suggest that inhibition of GRK2 function may constitute a new approach for treatment of flavivirus associated diseases. The Flavivirus genus includes several emergent and reemergent viruses, such as dengue and yellow fever viruses, which cause severe diseases in humans for which there is no approved treatment. Flaviviruses are transmitted to humans by arthropods and they rely on scores of vertebrate and invertebrate factors to replicate in these disparate hosts. Identifying the host factors involved in viral propagation is critical to understanding the molecular mechanisms of infection and the development of new therapeutics. To identify human host factors required for yellow fever virus propagation, we completed two genome-scale siRNA screens. Among the candidates discovered were the G protein-coupled receptor kinases GRK2 and GRK4. We focused on the protein GRK2, a kinase first identified for its role in cellular signal transduction. We found that GRK2 was a host factor needed for productive infection by yellow fever, dengue and hepatitis C viruses and was required for both viral entry and efficient replication of the viral genome. GRKs, which are considered druggable, may be used as targets to develop broadspectrum anti-flavivirals.
登录
查看更多内容
影响因子：
3.7
作者：
Clyde, Karen;Barrera, Julio;Harris, Eva
通讯作者：
Harris, Eva
影响因子：
14.8
作者：
Kato, Takanobu;Date, Tomoko;Wakita, Takaji
通讯作者：
Wakita, Takaji
DOI：
10.1073/pnas.83.9.2797
发表时间：
1986-05-01
影响因子：
11.1
作者：
BENOVIC, JL;STRASSER, RH;LEFKOWITZ, RJ
通讯作者：
LEFKOWITZ, RJ
影响因子：
64.8
作者：
Deng, HK;Liu, R;Landau, NR
通讯作者：
Landau, NR
影响因子：
64.8
作者：
Krishnan, Manoj N.;Ng, Aylwin;Sukumaran, Bindu;Gilfoy, Felicia D.;Uchil, Pradeep D.;Sultana, Hameeda;Brass, Abraham L.;Adametz, Rachel;Tsui, Melody;Qian, Feng;Montgomery, Ruth R.;Lev, Sima;Mason, Peter W.;Koski, Raymond A.;Elledge, Stephen J.;Xavier, Ramnik J.;Agaisse, Herve;Fikrig, Erol
通讯作者：
Fikrig, Erol