Capped RNA primer binding to influenza polymerase and implications for the mechanism of cap-binding inhibitors.
Capped RNA primer binding to influenza polymerase and implications for the mechanism of cap-binding inhibitors.
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RNA引物与流感聚合酶的结合及其对盖结合抑制剂机制的影响。
DOI：
10.1093/nar/gkx1210
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发表时间：
2018-01-25
影响因子：
14.9
通讯作者：
Cusack S
中科院分区：
文献类型：
--
作者：
Pflug A;Gaudon S;Resa-Infante P;Lethier M;Reich S;Schulze WM;Cusack S
流感聚合酶使用从新生 Pol II 转录物中抓取的短加帽引物来启动病毒 mRNA 的转录。在这里，我们描述了甲型和乙型流感聚合酶的晶体结构，其以与转录起始（“引发状态”）一致的配置与加帽引物结合，并通过功能测定表明，PB2中链和帽结合域的保守残基对于定位加帽RNA非常重要。特别是，PB2 Arg264 的突变与帽中的三磷酸键相互作用，显着且特异性地降低帽依赖性转录。我们还比较了启动状态下中链和帽结合结构域的配置以及它们在结合有效帽结合抑制剂 VX-787 或相近类似物的结构中非常不同的相对排列（称为“apo”状态）。在“apo”状态下，抑制剂与中链结构域产生额外的相互作用，从而增加了其亲和力，超出了单独与帽结合结构域的亲和力。比较表明，只有当 VX-787 具有双重作用模式（直接抑制加帽 RNA 结合以及稳定转录失活的“apo”状态）时，才能合理解释允许病毒逃离 VX-787（特别是 PB2 N510T）的某些突变的耐药机制。
Influenza polymerase uses short capped primers snatched from nascent Pol II transcripts to initiate transcription of viral mRNAs. Here we describe crystal structures of influenza A and B polymerase bound to a capped primer in a configuration consistent with transcription initiation (’priming state’) and show by functional assays that conserved residues from both the PB2 midlink and cap-binding domains are important for positioning the capped RNA. In particular, mutation of PB2 Arg264, which interacts with the triphosphate linkage in the cap, significantly and specifically decreases cap-dependent transcription. We also compare the configuration of the midlink and cap-binding domains in the priming state with their very different relative arrangement (called the ‘apo’ state) in structures where the potent cap-binding inhibitor VX-787, or a close analogue, is bound. In the ‘apo’ state the inhibitor makes additional interactions to the midlink domain that increases its affinity beyond that to the cap-binding domain alone. The comparison suggests that the mechanism of resistance of certain mutations that allow virus to escape from VX-787, notably PB2 N510T, can only be rationalized if VX-787 has a dual mode of action, direct inhibition of capped RNA binding as well as stabilization of the transcriptionally inactive ‘apo’ state.
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影响因子：
4.2
作者：
Bandarage, Upul K.;Clarke, Michael P.;Charifson, Paul S.
通讯作者：
Charifson, Paul S.
流感病毒聚合酶复合物：抗病毒药物设计的结构，功能和重要性的更新。
影响因子：
13.3
作者：
Stevaert, Annelies;Naesens, Lieve
通讯作者：
Naesens, Lieve
影响因子：
5.4
作者：
Poon, LLM;Pritlove, DC;Brownlee, GG
通讯作者：
Brownlee, GG
DOI：
10.1073/pnas.0507415102
发表时间：
2005-12-20
影响因子：
11.1
作者：
Gabriel, G;Dauber, B;Stech, J
通讯作者：
Stech, J
影响因子：
64.8
作者：
Hengrung N;El Omari K;Serna Martin I;Vreede FT;Cusack S;Rambo RP;Vonrhein C;Bricogne G;Stuart DI;Grimes JM;Fodor E
通讯作者：
Fodor E