Subgenomic flavivirus RNA binds the mosquito DEAD/H-box helicase ME31B and determines Zika virus transmission by Aedes aegypti
Subgenomic flavivirus RNA binds the mosquito DEAD/H-box helicase ME31B and determines Zika virus transmission by Aedes aegypti
复制标题
DOI：
10.1073/pnas.1905617116
复制
发表时间：
2019-09-17
影响因子：
11.1
通讯作者：
Pijlman, Gorben P.
中科院分区：
文献类型：
--
作者：
Goertz, Giel P.;van Bree, Joyce W. M.;Pijlman, Gorben P.
寨卡病毒（ZIKV）是一种节肢动物传播的黄病毒，主要由埃及伊蚊传播，对全球人类健康构成威胁。所有黄病毒，包括那些只在蚊子中复制的黄病毒，在感染细胞中产生大量的非编码亚基因组黄病毒RNA (sfRNA)，这表明sfRNA在蚊子感染过程中具有重要功能。目前，sfRNA在蚊子传播黄病毒中的作用尚不清楚。在这里，我们证明了缺乏sfrna的ZIKV （ZIKV Delta SF1）在蚊子细胞培养中复制与野生型ZIKV相似，但在伊蚊传播中被严重减弱。寨卡病毒三角洲SF1唾液阳性的蚊子占5%，寨卡病毒为31%。此外，与寨卡病毒相比，寨卡病毒δ SF1在蚊子唾液中的滴度较低。通过传染性血餐和胸内注射对蚊子感染的比较表明，sfRNA对寨卡病毒克服蚊子中肠屏障和促进病毒在唾液中积聚起重要作用。新一代测序结果显示，与寨卡病毒感染相比，寨卡病毒δ SF1感染后病毒小干扰rna升高。rna亲和纯化后的质谱分析发现，sfRNA特异性地与一组特定的Ae。通常与RNA周转和蛋白质翻译相关的埃及伊蚊蛋白质。DEAD/H-box解旋酶ME31B对sfRNA的亲和力最高，对sfRNA具有抗病毒活性。蚊子的寨卡病毒。蚊细胞。基于这些结果，我们提出了一个sfRNA隔离ME31B促进黄病毒复制和病毒粒子产生的机制模型，从而促进蚊子传播。
Zika virus (ZIKV) is an arthropod-borne flavivirus predominantly transmitted by Aedes aegypti mosquitoes and poses a global human health threat. All flaviviruses, including those that exclusively replicate in mosquitoes, produce a highly abundant, noncoding subgenomic flavivirus RNA (sfRNA) in infected cells, which implies an important function of sfRNA during mosquito infection. Currently, the role of sfRNA in flavivirus transmission by mosquitoes is not well understood. Here, we demonstrate that an sfRNA-deficient ZIKV (ZIKV Delta SF1) replicates similar to wild-type ZIKV in mosquito cell culture but is severely attenuated in transmission by Ae. aegypti after an infectious blood meal, with 5% saliva-positive mosquitoes for ZIKV Delta SF1 vs. 31% for ZIKV. Furthermore, viral titers in the mosquito saliva were lower for ZIKV Delta SF1 as compared to ZIKV. Comparison of mosquito infection via infectious blood meals and intrathoracic injections showed that sfRNA is important for ZIKV to overcome the mosquito midgut barrier and to promote virus accumulation in the saliva. Next-generation sequencing of infected mosquitoes showed that viral small-interfering RNAs were elevated upon ZIKV Delta SF1 as compared to ZIKV infection. RNA-affinity purification followed by mass spectrometry analysis uncovered that sfRNA specifically interacts with a specific set of Ae. aegypti proteins that are normally associated with RNA turnover and protein translation. The DEAD/H-box helicase ME31B showed the highest affinity for sfRNA and displayed antiviral activity against. ZIKV in Ae. aegypti cells. Based on these results, we present a mechanistic model in which sfRNA sequesters ME31B to promote flavivirus replication and virion production to facilitate transmission by mosquitoes.