PDlim2 selectively interacts with the PDZ binding motif of highly pathogenic avian H5N1 influenza A virus NS1.
PDlim2 selectively interacts with the PDZ binding motif of highly pathogenic avian H5N1 influenza A virus NS1.
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PDlim2 选择性地与高致病性禽 H5N1 甲型流感病毒 NS1 的 PDZ 结合基序相互作用
DOI：
10.1371/journal.pone.0019511
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发表时间：
2011
期刊：
影响因子：
3.7
通讯作者：
Cao Y
中科院分区：
文献类型：
--
作者：
Yu J;Li X;Wang Y;Li B;Li H;Li Y;Zhou W;Zhang C;Wang Y;Rao Z;Bartlam M;Cao Y
甲型流感病毒的多功能NS 1蛋白是一种病毒毒力决定因子。NS 1 C末端的最后四个残基构成I型PDZ结构域结合基序（PBM）。目前流行的禽流感病毒携带具有共有序列ESEV的NS 1 PBM，而人流感病毒携带具有共有序列RSKV或RSEV的NS 1 PBM。据报道，甲型流感病毒NS 1的PBM序列在动物研究中有助于高病毒致病性。本研究利用酵母双杂交技术筛选出PDlim 2作为高致病性禽流感病毒H5 N1株与ESEV（A/Chicken/Henan/12/2004/H5 N1，HN 12-NS 1）NS 1 PBM结合的新靶点。通过体外GST下拉试验以及体内哺乳动物双杂交试验和双分子荧光互补试验证实了这种相互作用。结合也被证实是介导的PDlim 2 PDZ结构域与NS 1 PBM基序的相互作用。有趣的是，我们的测定显示PDlim 2与HN 12-NS 1特异性结合，但与携带RSEV PBM的人流感H1 N1病毒（A/波多黎各/8/34/H1 N1，PR 8-NS 1）的NS 1没有结合。PDlim 2 PDZ结构域与来自HN 12-NS 1的C-末端六肽融合的晶体结构，连同对PDlim 2突变体的GST下拉测定，揭示了PDlim 2的残基Arg 16和Lys 31对于PDlim 2和HN 12-NS 1之间的结合是关键的。HN 12-NS 1（ESEV）而非PR 8-NS 1（RSEV）的选择性结合靶的鉴定使我们能够提出NS 1 PBM与PDlim 2或其他含PDZ蛋白之间相互作用的结构机制。
The multi-functional NS1 protein of influenza A virus is a viral virulence determining factor. The last four residues at the C-terminus of NS1 constitute a type I PDZ domain binding motif (PBM). Avian influenza viruses currently in circulation carry an NS1 PBM with consensus sequence ESEV, whereas human influenza viruses bear an NS1 PBM with consensus sequence RSKV or RSEV. The PBM sequence of the influenza A virus NS1 is reported to contribute to high viral pathogenicity in animal studies. Here, we report the identification of PDlim2 as a novel binding target of the highly pathogenic avian influenza virus H5N1 strain with an NS1 PBM of ESEV (A/Chicken/Henan/12/2004/H5N1, HN12-NS1) by yeast two-hybrid screening. The interaction was confirmed by in vitro GST pull-down assays, as well as by in vivo mammalian two-hybrid assays and bimolecular fluorescence complementation assays. The binding was also confirmed to be mediated by the interaction of the PDlim2 PDZ domain with the NS1 PBM motif. Interestingly, our assays showed that PDlim2 bound specifically with HN12-NS1, but exhibited no binding to NS1 from a human influenza H1N1 virus bearing an RSEV PBM (A/Puerto Rico/8/34/H1N1, PR8-NS1). A crystal structure of the PDlim2 PDZ domain fused with the C-terminal hexapeptide from HN12-NS1, together with GST pull-down assays on PDlim2 mutants, reveals that residues Arg16 and Lys31 of PDlim2 are critical for the binding between PDlim2 and HN12-NS1. The identification of a selective binding target of HN12-NS1 (ESEV), but not PR8-NS1 (RSEV), enables us to propose a structural mechanism for the interaction between NS1 PBM and PDlim2 or other PDZ-containing proteins.
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影响因子：
4.8
作者：
Katz A;Freiberg AN;Backström V;Schulz AR;Mateos A;Holm L;Pettersson RF;Vaheri A;Flick R;Plyusnin A
通讯作者：
Plyusnin A
DOI：
10.1107/s0907444904019158
发表时间：
2004-12-01
影响因子：
2.2
作者：
Emsley, P;Cowtan, K
通讯作者：
Cowtan, K
影响因子：
4.8
作者：
Karthikeyan, S;Leung, TL;Ladias, JAA
通讯作者：
Ladias, JAA
影响因子：
56.9
作者：
Songyang, Z;Fanning, AS;Cantley, LC
通讯作者：
Cantley, LC
影响因子：
82.9
作者：
Seo, SH;Hoffmann, E;Webster, RG
通讯作者：
Webster, RG