ID NS1_I34A1 Reviewed; 230 AA. AC P03496; Q20N32; Q67267; Q71QT3; DT 21-JUL-1986, integrated into UniProtKB/Swiss-Prot. DT 21-JUL-1986, sequence version 1. DT 24-JAN-2024, entry version 150. DE RecName: Full=Non-structural protein 1 {ECO:0000255|HAMAP-Rule:MF_04066}; DE Short=NS1 {ECO:0000255|HAMAP-Rule:MF_04066}; DE AltName: Full=NS1A {ECO:0000255|HAMAP-Rule:MF_04066}; GN Name=NS {ECO:0000255|HAMAP-Rule:MF_04066}; OS Influenza A virus (strain A/Puerto Rico/8/1934 H1N1). OC Viruses; Riboviria; Orthornavirae; Negarnaviricota; Polyploviricotina; OC Insthoviricetes; Articulavirales; Orthomyxoviridae; Alphainfluenzavirus; OC Alphainfluenzavirus influenzae; Influenza A virus. OX NCBI_TaxID=211044; OH NCBI_TaxID=8782; Aves. OH NCBI_TaxID=9606; Homo sapiens (Human). OH NCBI_TaxID=9823; Sus scrofa (Pig). RN [1] RP NUCLEOTIDE SEQUENCE [GENOMIC RNA]. RX PubMed=7465426; DOI=10.1093/nar/8.23.5845; RA Baez M., Taussig R., Zazra J.J., Young J.F., Palese P., Reisfeld A., RA Skalka A.M.; RT "Complete nucleotide sequence of the influenza A/PR/8/34 virus NS gene and RT comparison with the NS genes of the A/Udorn/72 and A/FPV/Rostock/34 RT strains."; RL Nucleic Acids Res. 8:5845-5858(1980). RN [2] RP NUCLEOTIDE SEQUENCE [GENOMIC RNA]. RX PubMed=11779399; DOI=10.1098/rstb.2001.0979; RA Schickli J.H., Flandorfer A., Nakaya T., Martinez-Sobrido L., RA Garcia-Sastre A., Palese P.; RT "Plasmid-only rescue of influenza A virus vaccine candidates."; RL Philos. Trans. R. Soc. Lond., B, Biol. Sci. 356:1965-1973(2001). RN [3] RP NUCLEOTIDE SEQUENCE [GENOMIC RNA], AND REVERSE GENETICS. RX PubMed=15163504; DOI=10.1016/j.virusres.2004.02.028; RA de Wit E., Spronken M.I.J., Bestebroer T.M., Rimmelzwaan G.F., RA Osterhaus A.D.M.E., Fouchier R.A.M.; RT "Efficient generation and growth of influenza virus A/PR/8/34 from eight RT cDNA fragments."; RL Virus Res. 103:155-161(2004). RN [4] RP NUCLEOTIDE SEQUENCE [GENOMIC RNA]. RA Ghedin E., Spiro D., Miller N., Zaborsky J., Feldblyum T., Subbu V., RA Shumway M., Sparenborg J., Groveman L., Halpin R., Sitz J., Koo H., RA Salzberg S.L., Webster R.G., Hoffmann E., Krauss S., Naeve C., Bao Y., RA Bolotov P., Dernovoy D., Kiryutin B., Lipman D.J., Tatusova T.; RT "The NIAID influenza genome sequencing project."; RL Submitted (MAR-2006) to the EMBL/GenBank/DDBJ databases. RN [5] RP REVIEW. RX PubMed=12758165; DOI=10.1016/s0042-6822(03)00119-3; RA Krug R.M., Yuan W., Noah D.L., Latham A.G.; RT "Intracellular warfare between human influenza viruses and human cells: the RT roles of the viral NS1 protein."; RL Virology 309:181-189(2003). RN [6] RP INTERACTION WITH HUMAN TRIM25, AND MUTAGENESIS OF ARG-38; LYS-41; GLU-96 RP AND GLU-97. RX PubMed=19454348; DOI=10.1016/j.chom.2009.04.006; RA Gack M.U., Albrecht R.A., Urano T., Inn K.-S., Huang I.-C., Carnero E., RA Farzan M., Inoue S., Jung J.U., Garcia-Sastre A.; RT "Influenza A virus NS1 targets the ubiquitin ligase TRIM25 to evade RT recognition by the host viral RNA sensor RIG-I."; RL Cell Host Microbe 5:439-449(2009). RN [7] RP ISGYLATION AT LYS-20; LYS-41; LYS-108; LYS-110; LYS-126; LYS-217 AND RP LYS-219, AND MUTAGENESIS OF LYS-20; LYS-41; LYS-108; LYS-110; LYS-126; RP LYS-217 AND LYS-219. RX PubMed=20385878; DOI=10.4049/jimmunol.0903588; RA Tang Y., Zhong G., Zhu L., Liu X., Shan Y., Feng H., Bu Z., Chen H., RA Wang C.; RT "Herc5 attenuates influenza A virus by catalyzing ISGylation of viral NS1 RT protein."; RL J. Immunol. 184:5777-5790(2010). RN [8] RP FUNCTION. RX PubMed=33766561; DOI=10.1016/j.jbc.2021.100579; RA Kesavardhana S., Samir P., Zheng M., Malireddi R.K.S., Karki R., RA Sharma B.R., Place D.E., Briard B., Vogel P., Kanneganti T.D.; RT "DDX3X coordinates host defense against influenza virus by activating the RT NLRP3 inflammasome and type I interferon response."; RL J. Biol. Chem. 296:100579-100579(2021). CC -!- FUNCTION: Inhibits post-transcriptional processing of cellular pre- CC mRNA, by binding and inhibiting two cellular proteins that are required CC for the 3'-end processing of cellular pre-mRNAs: the 30 kDa cleavage CC and polyadenylation specificity factor/CPSF4 and the poly(A)-binding CC protein 2/PABPN1. In turn, unprocessed 3' end pre-mRNAs accumulate in CC the host nucleus and are no longer exported to the cytoplasm. Cellular CC protein synthesis is thereby shut off very early after virus infection. CC Viral protein synthesis is not affected by the inhibition of the CC cellular 3' end processing machinery because the poly(A) tails of viral CC mRNAs are produced by the viral polymerase through a stuttering CC mechanism. Prevents the establishment of the cellular antiviral state CC by inhibiting TRIM25-mediated RIGI ubiquitination, which normally CC triggers the antiviral transduction signal that leads to the activation CC of type I IFN genes by transcription factors IRF3 and IRF7. Also binds CC poly(A) and U6 snRNA. Inhibits the integrated stress response (ISR) in CC the infected cell by blocking dsRNA binding by EIF2AK2/PKR and further CC phosphorylation of EIF2S1/EIF-2ALPHA (PubMed:33766561). Stress granule CC formation is thus inhibited, which allows protein synthesis and viral CC replication (PubMed:33766561). {ECO:0000255|HAMAP-Rule:MF_04066, CC ECO:0000269|PubMed:33766561}. CC -!- SUBUNIT: Homodimer. Interacts with host TRIM25 (via coiled coil); this CC interaction specifically inhibits TRIM25 multimerization and TRIM25- CC mediated RIGI CARD ubiquitination. Interacts with human EIF2AK2/PKR, CC CPSF4, IVNS1ABP and PABPN1. {ECO:0000255|HAMAP-Rule:MF_04066}. CC -!- INTERACTION: CC P03496; P03496: NS; NbExp=4; IntAct=EBI-2547442, EBI-2547442; CC P03496; P55265: ADAR; Xeno; NbExp=8; IntAct=EBI-2547442, EBI-2462104; CC P03496; Q12906: ILF3; Xeno; NbExp=3; IntAct=EBI-2547442, EBI-78756; CC P03496; P27986: PIK3R1; Xeno; NbExp=6; IntAct=EBI-2547442, EBI-79464; CC P03496; O00459: PIK3R2; Xeno; NbExp=13; IntAct=EBI-2547442, EBI-346930; CC P03496; O75569: PRKRA; Xeno; NbExp=3; IntAct=EBI-2547442, EBI-713955; CC P03496; O95793: STAU1; Xeno; NbExp=6; IntAct=EBI-2547442, EBI-358174; CC P03496; Q14258: TRIM25; Xeno; NbExp=3; IntAct=EBI-2547442, EBI-2341129; CC -!- SUBCELLULAR LOCATION: Host nucleus {ECO:0000255|HAMAP-Rule:MF_04066}. CC Host cytoplasm {ECO:0000255|HAMAP-Rule:MF_04066}. Note=In uninfected, CC transfected cells, NS1 is localized in the nucleus. Only in virus CC infected cells, the nuclear export signal is unveiled, presumably by a CC viral protein, and a fraction of NS1 is exported in the cytoplasm. CC {ECO:0000255|HAMAP-Rule:MF_04066}. CC -!- ALTERNATIVE PRODUCTS: CC Event=Alternative splicing; Named isoforms=2; CC Name=NS1; CC IsoId=P03496-1; Sequence=Displayed; CC Name=NEP; Synonyms=NS2; CC IsoId=P03508-1; Sequence=External; CC -!- DOMAIN: The dsRNA-binding region is required for suppression of RNA CC silencing. {ECO:0000255|HAMAP-Rule:MF_04066}. CC -!- PTM: Upon interferon induction, ISGylated via host HERC5; this results CC in the impairment of NS1 interaction with RNA targets due to its CC inability to form homodimers and to interact with host EIF2AK2/PKR. CC There are two ISGylated forms: one form is ISGylated at Lys-20, Lys-41, CC Lys-217, and Lys-219, and another one at Lys-108, Lys-110, and Lys-126, CC they represent band I and II respectively. Lys-126 and Lys-217 are CC critical for host antiviral response in vivo. {ECO:0000255|HAMAP- CC Rule:MF_04066, ECO:0000269|PubMed:20385878}. CC -!- SIMILARITY: Belongs to the influenza A viruses NS1 family. CC --------------------------------------------------------------------------- CC Copyrighted by the UniProt Consortium, see https://www.uniprot.org/terms CC Distributed under the Creative Commons Attribution (CC BY 4.0) License CC --------------------------------------------------------------------------- DR EMBL; V01104; CAA24292.1; ALT_SEQ; Genomic_RNA. DR EMBL; J02150; AAA43536.1; -; Genomic_RNA. DR EMBL; AF389122; AAM75163.1; -; Genomic_RNA. DR EMBL; EF467817; ABO21703.1; -; Genomic_RNA. DR EMBL; CY009448; ABD77680.1; -; Genomic_RNA. DR RefSeq; NP_040984.1; NC_002020.1. DR PDB; 2GX9; X-ray; 2.10 A; A/B=79-207. DR PDB; 2ZKO; X-ray; 1.70 A; A/B=1-70. DR PDB; 3L4Q; X-ray; 2.30 A; A/B=73-230. DR PDB; 3O9Q; X-ray; 2.50 A; A/B=79-230. DR PDB; 3O9R; X-ray; 2.00 A; A/B=79-230. DR PDB; 3O9S; X-ray; 2.48 A; A/B=79-230. DR PDB; 3O9T; X-ray; 2.20 A; A/B=79-230. DR PDB; 3O9U; X-ray; 3.20 A; A/B/C/D/E/F/G/H=79-230. DR PDB; 3RVC; X-ray; 1.80 A; A=79-230. DR PDB; 5NT1; X-ray; 2.82 A; B/F/J=80-230. DR PDB; 5NT2; X-ray; 4.26 A; C/D/E/F=1-230. DR PDBsum; 2GX9; -. DR PDBsum; 2ZKO; -. DR PDBsum; 3L4Q; -. DR PDBsum; 3O9Q; -. DR PDBsum; 3O9R; -. DR PDBsum; 3O9S; -. DR PDBsum; 3O9T; -. DR PDBsum; 3O9U; -. DR PDBsum; 3RVC; -. DR PDBsum; 5NT1; -. DR PDBsum; 5NT2; -. DR SMR; P03496; -. DR DIP; DIP-29081N; -. DR IntAct; P03496; 222. DR MINT; P03496; -. DR ChEMBL; CHEMBL4523164; -. DR GeneID; 956533; -. DR KEGG; vg:956533; -. DR OrthoDB; 8721at10239; -. DR Reactome; R-HSA-1169408; ISG15 antiviral mechanism. DR Reactome; R-HSA-168276; NS1 Mediated Effects on Host Pathways. DR Reactome; R-HSA-168315; Inhibition of Host mRNA Processing and RNA Silencing. DR Reactome; R-HSA-168888; Inhibition of IFN-beta. DR Reactome; R-HSA-169131; Inhibition of PKR. DR Reactome; R-HSA-192823; Viral mRNA Translation. DR Reactome; R-HSA-5213460; RIPK1-mediated regulated necrosis. DR Reactome; R-HSA-9686347; Microbial modulation of RIPK1-mediated regulated necrosis. DR Reactome; R-HSA-9833482; PKR-mediated signaling. DR EvolutionaryTrace; P03496; -. DR Proteomes; UP000009255; Genome. DR Proteomes; UP000116373; Genome. DR Proteomes; UP000170967; Genome. DR GO; GO:0030430; C:host cell cytoplasm; IEA:UniProtKB-SubCell. DR GO; GO:0042025; C:host cell nucleus; IEA:UniProtKB-SubCell. DR GO; GO:0042802; F:identical protein binding; IPI:IntAct. DR GO; GO:0030291; F:protein serine/threonine kinase inhibitor activity; IEA:UniProtKB-KW. DR GO; GO:0003723; F:RNA binding; IEA:UniProtKB-KW. DR GO; GO:0039524; P:suppression by virus of host mRNA processing; IEA:UniProtKB-KW. DR GO; GO:0039580; P:suppression by virus of host PKR signaling; IEA:UniProtKB-KW. DR GO; GO:0039502; P:suppression by virus of host type I interferon-mediated signaling pathway; IEA:UniProtKB-KW. DR GO; GO:0039540; P:suppression by virus of host viral-induced cytoplasmic pattern recognition receptor signaling pathway via inhibition of host RIG-I activity; IEA:UniProtKB-KW. DR Gene3D; 3.30.420.330; Influenza virus non-structural protein, effector domain; 1. DR Gene3D; 1.10.287.10; S15/NS1, RNA-binding; 1. DR HAMAP; MF_04066; INFV_NS1; 1. DR InterPro; IPR004208; NS1. DR InterPro; IPR000256; NS1A. DR InterPro; IPR038064; NS1A_effect_dom-like_sf. DR InterPro; IPR009068; uS15_NS1_RNA-bd_sf. DR Pfam; PF00600; Flu_NS1; 1. DR SUPFAM; SSF143021; Ns1 effector domain-like; 1. DR SUPFAM; SSF47060; S15/NS1 RNA-binding domain; 1. PE 1: Evidence at protein level; KW 3D-structure; Alternative splicing; KW Eukaryotic host gene expression shutoff by virus; Host cytoplasm; KW Host gene expression shutoff by virus; Host mRNA suppression by virus; KW Host nucleus; Host-virus interaction; KW Inhibition of host innate immune response by virus; KW Inhibition of host interferon signaling pathway by virus; KW Inhibition of host PKR by virus; KW Inhibition of host pre-mRNA processing by virus; KW Inhibition of host RIG-I by virus; Inhibition of host RLR pathway by virus; KW Interferon antiviral system evasion; Isopeptide bond; Reference proteome; KW RNA-binding; Ubl conjugation; Viral immunoevasion. FT CHAIN 1..230 FT /note="Non-structural protein 1" FT /id="PRO_0000078945" FT REGION 1..73 FT /note="RNA-binding and homodimerization" FT /evidence="ECO:0000255|HAMAP-Rule:MF_04066" FT REGION 180..215 FT /note="CPSF4-binding" FT /evidence="ECO:0000255|HAMAP-Rule:MF_04066" FT REGION 205..230 FT /note="Disordered" FT /evidence="ECO:0000256|SAM:MobiDB-lite" FT REGION 223..230 FT /note="PABPN1-binding" FT /evidence="ECO:0000255|HAMAP-Rule:MF_04066" FT MOTIF 34..38 FT /note="Nuclear localization signal" FT /evidence="ECO:0000255|HAMAP-Rule:MF_04066" FT MOTIF 137..146 FT /note="Nuclear export signal" FT /evidence="ECO:0000255|HAMAP-Rule:MF_04066" FT CROSSLNK 20 FT /note="Glycyl lysine isopeptide (Lys-Gly) (interchain with FT G-Cter in ISG15); in band I form; by host" FT /evidence="ECO:0000269|PubMed:20385878" FT CROSSLNK 41 FT /note="Glycyl lysine isopeptide (Lys-Gly) (interchain with FT G-Cter in ISG15); in band I form; by host" FT /evidence="ECO:0000269|PubMed:20385878" FT CROSSLNK 108 FT /note="Glycyl lysine isopeptide (Lys-Gly) (interchain with FT G-Cter in ISG15); in band II form; by host" FT /evidence="ECO:0000269|PubMed:20385878" FT CROSSLNK 110 FT /note="Glycyl lysine isopeptide (Lys-Gly) (interchain with FT G-Cter in ISG15); in band II form; by host" FT /evidence="ECO:0000269|PubMed:20385878" FT CROSSLNK 126 FT /note="Glycyl lysine isopeptide (Lys-Gly) (interchain with FT G-Cter in ISG15); in band II form; by host" FT /evidence="ECO:0000269|PubMed:20385878" FT CROSSLNK 217 FT /note="Glycyl lysine isopeptide (Lys-Gly) (interchain with FT G-Cter in ISG15); in band I form; by host" FT /evidence="ECO:0000269|PubMed:20385878" FT CROSSLNK 219 FT /note="Glycyl lysine isopeptide (Lys-Gly) (interchain with FT G-Cter in ISG15); in band I form; by host" FT /evidence="ECO:0000269|PubMed:20385878" FT MUTAGEN 20 FT /note="K->A: No of ISGylation of band I form; when FT associated with K-41; K-217 and K-219." FT /evidence="ECO:0000269|PubMed:20385878" FT MUTAGEN 38 FT /note="R->A: Complete loss of inhibition of RIGICARD FT ubiquitination; when associated with A-41." FT /evidence="ECO:0000269|PubMed:19454348" FT MUTAGEN 41 FT /note="K->A: Complete loss of inhibition of RIGICARD FT ubiquitination; when associated with A-38." FT /evidence="ECO:0000269|PubMed:19454348, FT ECO:0000269|PubMed:20385878" FT MUTAGEN 41 FT /note="K->A: No of ISGylation of band I form; when FT associated with K-20; K-217 and K-219." FT /evidence="ECO:0000269|PubMed:19454348, FT ECO:0000269|PubMed:20385878" FT MUTAGEN 96 FT /note="E->A: Complete loss of inhibition of RIGICARD FT ubiquitination; when associated with A-97." FT /evidence="ECO:0000269|PubMed:19454348" FT MUTAGEN 97 FT /note="E->A: Complete loss of inhibition of RIGICARD FT ubiquitination; when associated with A-96." FT /evidence="ECO:0000269|PubMed:19454348" FT MUTAGEN 108 FT /note="K->A: No of ISGylation of band II form; when FT associated with K-110 and K-126." FT /evidence="ECO:0000269|PubMed:20385878" FT MUTAGEN 110 FT /note="K->A: No of ISGylation of band II form; when FT associated with K-108 and K-126." FT /evidence="ECO:0000269|PubMed:20385878" FT MUTAGEN 126 FT /note="K->A: No of ISGylation of band II form; when FT associated with K-108 and K-110." FT /evidence="ECO:0000269|PubMed:20385878" FT MUTAGEN 217 FT /note="K->A: No of ISGylation of band I form; when FT associated with K-20; K-41 and K-219." FT /evidence="ECO:0000269|PubMed:20385878" FT MUTAGEN 219 FT /note="K->A: No of ISGylation of band I form; when FT associated with K-20; K-41 and K-217." FT /evidence="ECO:0000269|PubMed:20385878" FT CONFLICT 55 FT /note="E -> K (in Ref. 4; ABD77680)" FT CONFLICT 101 FT /note="D -> E (in Ref. 2; AAA43536)" FT HELIX 3..24 FT /evidence="ECO:0007829|PDB:2ZKO" FT HELIX 30..50 FT /evidence="ECO:0007829|PDB:2ZKO" FT HELIX 54..69 FT /evidence="ECO:0007829|PDB:2ZKO" FT STRAND 88..93 FT /evidence="ECO:0007829|PDB:3RVC" FT HELIX 95..99 FT /evidence="ECO:0007829|PDB:3RVC" FT STRAND 105..112 FT /evidence="ECO:0007829|PDB:3RVC" FT STRAND 115..120 FT /evidence="ECO:0007829|PDB:3RVC" FT STRAND 127..137 FT /evidence="ECO:0007829|PDB:3RVC" FT STRAND 140..151 FT /evidence="ECO:0007829|PDB:3RVC" FT STRAND 156..162 FT /evidence="ECO:0007829|PDB:3RVC" FT HELIX 171..187 FT /evidence="ECO:0007829|PDB:3RVC" FT STRAND 191..194 FT /evidence="ECO:0007829|PDB:3RVC" FT HELIX 196..201 FT /evidence="ECO:0007829|PDB:3RVC" SQ SEQUENCE 230 AA; 25868 MW; 2F7EC18E3EE7A4BE CRC64; MDPNTVSSFQ VDCFLWHVRK RVADQELGDA PFLDRLRRDQ KSLRGRGSTL GLDIETATRA GKQIVERILK EESDEALKMT MASVPASRYL TDMTLEEMSR DWSMLIPKQK VAGPLCIRMD QAIMDKNIIL KANFSVIFDR LETLILLRAF TEEGAIVGEI SPLPSLPGHT AEDVKNAVGV LIGGLEWNDN TVRVSETLQR FAWRSSNENG RPPLTPKQKR EMAGTIRSEV //