6o01

From Proteopedia

X-ray structure of H5N1-NS1 R38A K41A G71E mutant

Structural highlights

6o01 is a 1 chain structure with sequence from Influenza A virus (A/Vietnam/1196/2004(H5N1)). Full crystallographic information is available from OCA. For a guided tour on the structure components use FirstGlance.
Method:	X-ray diffraction, Resolution 3Å
Resources:	FirstGlance, OCA, PDBe, RCSB, PDBsum, ProSAT

Function

Q6QT26_9INFA Inhibits post-transcriptional processing of cellular pre-mRNA, by binding and inhibiting two cellular proteins that are required for the 3'-end processing of cellular pre-mRNAs: the 30 kDa cleavage and polyadenylation specificity factor/CPSF4 and the poly(A)-binding protein 2/PABPN1. In turn, unprocessed 3' end pre-mRNAs accumulate in the host nucleus and are no longer exported to the cytoplasm. Cellular protein synthesis is thereby shut off very early after virus infection. Viral protein synthesis is not affected by the inhibition of the cellular 3' end processing machinery because the poly(A) tails of viral mRNAs are produced by the viral polymerase through a stuttering mechanism.[RuleBase:RU362113][SAAS:SAAS01036581] Prevents the establishment of the cellular antiviral state by inhibiting TRIM25-mediated DDX58 ubiquitination, which normally triggers the antiviral transduction signal that leads to the activation of type I IFN genes by transcription factors IRF3 and IRF7. Prevents human EIF2AK2/PKR activation, either by binding double-strand RNA, or by interacting directly with EIF2AK2/PKR. This function may be important at the very beginning of the infection, when NS1 is mainly present in the cytoplasm. Also binds poly(A) and U6 snRNA.[RuleBase:RU362113][SAAS:SAAS01036591]

Publication Abstract from PubMed

Influenza A virus (IAV) non-structural protein 1 (NS1), a potent antagonist of host immune response, is capable of interacting with RNA and a wide range of cellular proteins. NS1 consists of an RNA-binding domain (RBD) and an effector domain (ED) separated by a flexible linker region (LR). H5N1-NS1 has a characteristic 5-residue deletion in the LR with either G (minor group) or E (major group) at the 71(st) position, and non-H5N1-NS1 contains E71 with an intact linker. Based on the orientation of ED with respect to RBD, previous crystallographic studies have shown that minor group H5N1-NS1(G71), a non-H5N1-NS1 (H6N6-NS1(E71)), and the LR-deletion mutant H6N6-NS1(Delta80-84/E71) mimicking the major group H5N1-NS1, exhibit 'open', 'semi-open', and 'closed' conformations, respectively, suggesting that NS1 exhibits strain-dependent conformational preference. Here we report the first crystal structure of a naturally occurring H5N1-NS1(E71) and show that it adopts an 'open' conformation similar to the minor group of H5N1-NS1 (H5N1-NS1(G71)). We also show that H6N6-NS1(Delta80-84/E71) under a different crystallization condition and H6N6-NS1(Delta80-84/G71) also exhibit 'open' conformations, suggesting NS1 can adopt an 'open' conformation irrespective of E or G at the 71(st) position. Our single-molecule FRET analysis to investigate the conformational preference of NS1 in solution showed that all NS1 constructs predominantly exist in 'open' conformation. Further, our co-immunoprecipitation and binding studies showed that they all bind to cellular factors with similar affinity. Taken together, our studies suggest that NS1 exhibits strain-independent structural plasticity that allows it to interact with a wide variety of cellular ligands during viral infection.IMPORTANCEIAV is responsible for several pandemics over the last century and continues to infect millions annually. The frequent rise in drug-resistant strains necessitates exploring novel targets for developing antiviral drugs that can reduce the global burden of influenza infection. Because of its critical role in the replication and pathogenesis of IAV, non-structural protein 1 (NS1) is a potential target for developing antivirals. Previous studies suggested that NS1 adopts strain-dependent 'open', 'semi-open', and 'closed' conformations. Here we show, based on three crystal structures, that NS1 irrespective of strain differences can adopt 'open' conformation. We further show that NS1 from different strains primarily exists in 'open' conformation in solution and binds to cellular proteins with similar affinity. Together, our findings suggest that conformational polymorphism facilitated by a flexible linker is intrinsic to NS1, and this may be the underlying factor allowing NS1 to bind several cellular factors during IAV replication.

Influenza A virus protein NS1 exhibits strain-independent conformational plasticity.,Mitra S, Kumar D, Hu L, Sankaran B, Moosa MM, Rice AP, Ferreon JC, Ferreon ACM, Prasad BVV J Virol. 2019 Aug 2. pii: JVI.00917-19. doi: 10.1128/JVI.00917-19. PMID:31375595^[1]

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.

References

↑ Mitra S, Kumar D, Hu L, Sankaran B, Moosa MM, Rice AP, Ferreon JC, Ferreon ACM, Prasad BVV. Influenza A virus protein NS1 exhibits strain-independent conformational plasticity. J Virol. 2019 Aug 2. pii: JVI.00917-19. doi: 10.1128/JVI.00917-19. PMID:31375595 doi:http://dx.doi.org/10.1128/JVI.00917-19