Preference of RIG-I for short viral RNA molecules in infected cells revealed by next-generation sequencing
Preference of RIG-I for short viral RNA molecules in infected cells revealed by next-generation sequencing
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DOI：
10.1073/pnas.1005077107
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发表时间：
2010-09-14
影响因子：
11.1
通讯作者：
Garcia-Sastre, Adolfo
中科院分区：
文献类型：
--
作者：
Baum, Alina;Sachidanandam, Ravi;Garcia-Sastre, Adolfo
病毒感染的细胞内检测是由称为病原体识别受体（PRR）的分子进行的先天免疫的关键组成部分。PRR通过其各自的病原体相关分子模式（PAMP）激活导致促炎细胞因子（包括I型IFN）的产生，并在宿主中建立抗病毒状态。在迄今为止发现的所有PRR中，已显示视黄酸诱导基因I（RIG-I）在RNA病毒识别中起关键作用。在体外和转染研究的基础上，在病毒复制过程中产生的5 'ppp RNA被认为是结合和激活这个重要的传感器。然而，在病毒感染过程中与内源性RIG-I相互作用的RNA分子的性质尚未确定。在这项工作中，我们使用下一代RNA测序显示，RIG-I优先与较短的，5 'ppp在感染细胞中含有病毒RNA分子。我们发现，在仙台感染期间，RIG-I特异性结合缺陷干扰（DI）颗粒的基因组，而不结合全长病毒基因组或任何其他病毒RNA。在流感感染的细胞中，RIG-I优先与较短的基因组片段以及亚基因组DI颗粒相关。我们的分析第一次确定RIG-I PAMPs在自然感染条件下，并意味着全长基因组的单节段RNA病毒家族在感染过程中不受RIG-I的约束。
Intracellular detection of virus infections is a critical component of innate immunity carried out by molecules known as pathogen recognition receptors (PRRs). Activation of PRRs by their respective pathogen-associated molecular patterns (PAMPs) leads to production of proinflamatory cytokines, including type I IFN, and the establishment of an antiviral state in the host. Out of all PRRs found to date, retinoic acid inducible gene I (RIG-I) has been shown to play a key role in recognition of RNA viruses. On the basis of in vitro and transfection studies, 5'ppp RNA produced during virus replication is thought to bind and activate this important sensor. However, the nature of RNA molecules that interact with endogenous RIG-I during the course of viral infection has not been determined. In this work we use next-generation RNA sequencing to show that RIG-I preferentially associates with shorter, 5'ppp containing viral RNA molecules in infected cells. We found that during Sendai infection RIG-I specifically bound the genome of the defective interfering (DI) particle and did not bind the full-length virus genome or any other viral RNAs. In influenza-infected cells RIG-I preferentially associated with shorter genomic segments as well as subgenomic DI particles. Our analysis for the first time identifies RIG-I PAMPs under natural infection conditions and implies that full-length genomes of single segmented RNA virus families are not bound by RIG-I during infection.