POLN_EEVVT
ID POLN_EEVVT Reviewed; 2493 AA.
AC P27282; A0A0M3T9D8; O90163; Q5XQC5; Q66592; Q66594;
DT 01-AUG-1992, integrated into UniProtKB/Swiss-Prot.
DT 10-APR-2019, sequence version 3.
DT 03-AUG-2022, entry version 161.
DE RecName: Full=Polyprotein P1234;
DE Short=P1234;
DE AltName: Full=Non-structural polyprotein;
DE Contains:
DE RecName: Full=Polyprotein P123';
DE Short=P123';
DE Contains:
DE RecName: Full=Polyprotein P123;
DE Short=P123;
DE Contains:
DE RecName: Full=mRNA-capping enzyme nsP1;
DE EC=2.1.1.- {ECO:0000269|PubMed:26041283};
DE EC=2.7.7.- {ECO:0000269|PubMed:26041283};
DE AltName: Full=Non-structural protein 1;
DE Contains:
DE RecName: Full=Protease nsP2;
DE EC=3.1.3.33 {ECO:0000250|UniProtKB:P08411};
DE EC=3.4.22.- {ECO:0000250|UniProtKB:Q8JUX6};
DE EC=3.6.1.15 {ECO:0000250|UniProtKB:Q8JUX6};
DE EC=3.6.4.13 {ECO:0000250|UniProtKB:Q8JUX6};
DE AltName: Full=Non-structural protein 2;
DE Short=nsP2;
DE Contains:
DE RecName: Full=Non-structural protein 3';
DE Short=nsP3';
DE EC=3.1.3.84 {ECO:0000305};
DE Contains:
DE RecName: Full=Non-structural protein 3;
DE Short=nsP3;
DE EC=3.1.3.84 {ECO:0000269|PubMed:27440879};
DE Contains:
DE RecName: Full=RNA-directed RNA polymerase nsP4;
DE EC=2.7.7.19 {ECO:0000250|UniProtKB:P03317};
DE EC=2.7.7.48 {ECO:0000255|PROSITE-ProRule:PRU00539};
DE AltName: Full=Non-structural protein 4;
DE Short=nsP4;
OS Venezuelan equine encephalitis virus (strain Trinidad donkey) (VEEV).
OC Viruses; Riboviria; Orthornavirae; Kitrinoviricota; Alsuviricetes;
OC Martellivirales; Togaviridae; Alphavirus.
OX NCBI_TaxID=11038;
OH NCBI_TaxID=9913; Bos taurus (Bovine).
OH NCBI_TaxID=9268; Didelphis marsupialis (Southern opossum).
OH NCBI_TaxID=9793; Equus asinus (Donkey) (Equus africanus asinus).
OH NCBI_TaxID=9796; Equus caballus (Horse).
OH NCBI_TaxID=9606; Homo sapiens (Human).
OH NCBI_TaxID=53535; Melanoconion.
OH NCBI_TaxID=9272; Philander opossum (Gray four-eyed opossum).
OH NCBI_TaxID=10162; Proechimys.
OH NCBI_TaxID=42415; Sigmodon hispidus (Hispid cotton rat).
RN [1]
RP NUCLEOTIDE SEQUENCE [GENOMIC RNA].
RC STRAIN=TC-83, and Trinidad donkey;
RX PubMed=2524126; DOI=10.1016/0042-6822(89)90347-4;
RA Kinney R.M., Johnson B.J.B., Welch J.B., Tsuchiya K.R., Trent D.W.;
RT "The full-length nucleotide sequences of the virulent Trinidad donkey
RT strain of Venezuelan equine encephalitis virus and its attenuated vaccine
RT derivative, strain TC-83.";
RL Virology 170:19-30(1989).
RN [2]
RP SEQUENCE REVISION.
RA Kinney R.;
RL Submitted (MAY-1999) to the EMBL/GenBank/DDBJ databases.
RN [3]
RP NUCLEOTIDE SEQUENCE [GENOMIC RNA].
RC STRAIN=71-180;
RX PubMed=1469368; DOI=10.1099/0022-1317-73-12-3301;
RA Kinney R.M., Tsuchiya K.R., Sneider J.M., Trent D.W.;
RT "Molecular evidence for the origin of the widespread Venezuelan equine
RT encephalitis epizootic of 1969 to 1972.";
RL J. Gen. Virol. 73:3301-3305(1992).
RN [4]
RP NUCLEOTIDE SEQUENCE [GENOMIC RNA].
RC STRAIN=V3526;
RA Davis N., Johnston R.E., Smith J.F., Crise B., Parker M.D.;
RT "Venezuelan equine encephalitis virus strain V3526.";
RL Submitted (SEP-2004) to the EMBL/GenBank/DDBJ databases.
RN [5]
RP NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
RC STRAIN=COAN5506 {ECO:0000312|EMBL:ALE15112.1};
RG Consortium for Microbial Forensics and Genomics (microFORGE);
RA Forrester N.L., Auguste A.J., Guerbois M., Rossi S.L., Lin D., Hari K.,
RA Weaver S.C.;
RT "Draft genome sequence of Venezuelan equine encephalitis virus strain
RT CoAn5506, a 1967 equine isolate from Colombia.";
RL Submitted (APR-2015) to the EMBL/GenBank/DDBJ databases.
RN [6]
RP SUBCELLULAR LOCATION (PROTEASE NSP2), NUCLEAR LOCALIZATION SIGNAL,
RP INTERACTION WITH HOST KPNA1 (PROTEASE NSP2), NUCLEAR EXPORT SIGNAL, DOMAIN
RP (PROTEASE NSP2), AND MUTAGENESIS OF VAL-1056; LEU-1061; LEU-1063 AND
RP LYS-1181.
RC STRAIN=pV3000;
RX PubMed=17652399; DOI=10.1128/jvi.00371-07;
RA Montgomery S.A., Johnston R.E.;
RT "Nuclear import and export of Venezuelan equine encephalitis virus
RT nonstructural protein 2.";
RL J. Virol. 81:10268-10279(2007).
RN [7]
RP MUTAGENESIS OF VAL-538; THR-540 AND GLN-1006.
RX PubMed=23365438; DOI=10.1128/jvi.03142-12;
RA Kim D.Y., Atasheva S., Frolova E.I., Frolov I.;
RT "Venezuelan equine encephalitis virus nsP2 protein regulates packaging of
RT the viral genome into infectious virions.";
RL J. Virol. 87:4202-4213(2013).
RN [8]
RP CATALYTIC ACTIVITY (MRNA-CAPPING ENZYME NSP1), MUTAGENESIS OF HIS-37;
RP HIS-45; ASP-63; GLU-118; TYR-285; ASP-354; ARG-365; ASN-369 AND ASN-375,
RP FUNCTION (MRNA-CAPPING ENZYME NSP1), ACTIVITY REGULATION (MRNA-CAPPING
RP ENZYME NSP1), AND COFACTOR (MRNA-CAPPING ENZYME NSP1).
RX PubMed=26041283; DOI=10.1128/jvi.00599-15;
RA Li C., Guillen J., Rabah N., Blanjoie A., Debart F., Vasseur J.J.,
RA Canard B., Decroly E., Coutard B.;
RT "mRNA capping by venezuelan equine encephalitis virus nsp1: functional
RT characterization and implications for antiviral research.";
RL J. Virol. 89:8292-8303(2015).
RN [9]
RP INTERACTION WITH HOST DDX1 (NON-STRUCTURAL PROTEIN 3), AND INTERACTION WITH
RP HOST DDX3 (NON-STRUCTURAL PROTEIN 3).
RX PubMed=27105836; DOI=10.1016/j.antiviral.2016.04.008;
RA Amaya M., Brooks-Faulconer T., Lark T., Keck F., Bailey C., Raman V.,
RA Narayanan A.;
RT "Venezuelan equine encephalitis virus non-structural protein 3 (nsP3)
RT interacts with RNA helicases DDX1 and DDX3 in infected cells.";
RL Antiviral Res. 131:49-60(2016).
RN [10]
RP FUNCTION (PROTEASE NSP2), AND MUTAGENESIS OF PRO-1248 AND GLN-1274.
RX PubMed=27318152; DOI=10.1016/j.virol.2016.06.005;
RA Bhalla N., Sun C., Metthew Lam L.K., Gardner C.L., Ryman K.D.,
RA Klimstra W.B.;
RT "Host translation shutoff mediated by non-structural protein 2 is a
RT critical factor in the antiviral state resistance of Venezuelan equine
RT encephalitis virus.";
RL Virology 496:147-165(2016).
RN [11]
RP FUNCTION (NON-STRUCTURAL PROTEIN 3), DOMAIN (NON-STRUCTURAL PROTEIN 3),
RP INTERACTION WITH HOST FXR1 (NON-STRUCTURAL PROTEIN 3), INTERACTION WITH
RP HOST FXR2 (NON-STRUCTURAL PROTEIN 3), AND INTERACTION WITH HOST FMR1
RP (NON-STRUCTURAL PROTEIN 3).
RX PubMed=27509095; DOI=10.1371/journal.ppat.1005810;
RA Kim D.Y., Reynaud J.M., Rasalouskaya A., Akhrymuk I., Mobley J.A.,
RA Frolov I., Frolova E.I.;
RT "New World and Old World Alphaviruses Have Evolved to Exploit Different
RT Components of Stress Granules, FXR and G3BP Proteins, for Assembly of Viral
RT Replication Complexes.";
RL PLoS Pathog. 12:E1005810-E1005810(2016).
RN [12]
RP DOMAIN (NON-STRUCTURAL PROTEIN 3), CATALYTIC ACTIVITY (NON-STRUCTURAL
RP PROTEIN 3), AND FUNCTION (NON-STRUCTURAL PROTEIN 3).
RX PubMed=27440879; DOI=10.1128/jvi.00705-16;
RA Li C., Debing Y., Jankevicius G., Neyts J., Ahel I., Coutard B., Canard B.;
RT "Viral Macro Domains Reverse Protein ADP-Ribosylation.";
RL J. Virol. 90:8478-8486(2016).
RN [13]
RP FUNCTION (NON-STRUCTURAL PROTEIN 3), DOMAIN (NON-STRUCTURAL PROTEIN 3), AND
RP CATALYTIC ACTIVITY (NON-STRUCTURAL PROTEIN 3).
RX PubMed=28150709; DOI=10.1038/srep41746;
RA Eckei L., Krieg S., Buetepage M., Lehmann A., Gross A., Lippok B.,
RA Grimm A.R., Kuemmerer B.M., Rossetti G., Luescher B., Verheugd P.;
RT "The conserved macrodomains of the non-structural proteins of Chikungunya
RT virus and other pathogenic positive strand RNA viruses function as mono-
RT ADP-ribosylhydrolases.";
RL Sci. Rep. 7:41746-41746(2017).
RN [14]
RP X-RAY CRYSTALLOGRAPHY (2.45 ANGSTROMS) OF 1003-1322, AND ACTIVE SITE
RP (PROTEASE NSP2).
RX PubMed=16962975; DOI=10.1016/j.str.2006.07.010;
RA Russo A.T., White M.A., Watowich S.J.;
RT "The crystal structure of the Venezuelan equine encephalitis alphavirus
RT nsP2 protease.";
RL Structure 14:1449-1458(2006).
RN [15] {ECO:0007744|PDB:5EZQ, ECO:0007744|PDB:5EZS}
RP X-RAY CRYSTALLOGRAPHY (1.66 ANGSTROMS) OF 992-1327, BIOPHYSICOCHEMICAL
RP PROPERTIES (PROTEASE NSP2), MUTAGENESIS OF ASN-1010; LYS-1015 AND ARG-1197,
RP AND CATALYTIC ACTIVITY (PROTASE NSP2).
RX PubMed=27030368; DOI=10.1021/acs.biochem.5b00992;
RA Hu X., Compton J.R., Leary D.H., Olson M.A., Lee M.S., Cheung J., Ye W.,
RA Ferrer M., Southall N., Jadhav A., Morazzani E.M., Glass P.J., Marugan J.,
RA Legler P.M.;
RT "Kinetic, Mutational, and Structural Studies of the Venezuelan Equine
RT Encephalitis Virus Nonstructural Protein 2 Cysteine Protease.";
RL Biochemistry 55:3007-3019(2016).
RN [16]
RP X-RAY CRYSTALLOGRAPHY (2.10 ANGSTROMS) OF 1003-1338, AND MUTAGENESIS OF
RP ASN-1010.
RX PubMed=29064679; DOI=10.1021/acs.biochem.7b00746;
RA Compton J.R., Mickey M.J., Hu X., Marugan J.J., Legler P.M.;
RT "Mutation of Asn-475 in the Venezuelan equine encephalitis virus nsP2
RT cysteine protease leads to a self-inhibited state.";
RL Biochemistry 56:6221-6230(2017).
CC -!- FUNCTION: [Polyprotein P1234]: Inactive precursor of the viral
CC replicase, which is activated by cleavages carried out by the viral
CC protease nsP2. {ECO:0000250|UniProtKB:Q8JUX6}.
CC -!- FUNCTION: [Polyprotein P123]: The early replication complex formed by
CC the polyprotein P123 and nsP4 synthesizes the minus-strand RNAs
CC (antigenome) (By similarity). Polyprotein P123 is a short-lived
CC polyprotein that accumulates during early stage of infection
CC (Probable). As soon P123 is cleaved into mature proteins, the plus-
CC strand RNAs synthesis begins (By similarity).
CC {ECO:0000250|UniProtKB:P03317, ECO:0000305}.
CC -!- FUNCTION: [Polyprotein P123']: The early replication complex formed by
CC the polyprotein P123' and nsP4 synthesizes minus-strand RNAs
CC (antigenome) (Probable). Polyprotein P123' is a short-lived polyprotein
CC that accumulates during early stage of infection (Probable). As soon
CC P123' is cleaved into mature proteins, the plus-strand RNAs synthesis
CC begins (Probable). {ECO:0000305}.
CC -!- FUNCTION: [mRNA-capping enzyme nsP1]: Cytoplasmic capping enzyme that
CC catalyzes two virus-specific reactions: methyltransferase and nsP1
CC guanylyltransferase (PubMed:26041283). mRNA-capping is necessary since
CC all viral RNAs are synthesized in the cytoplasm, and host capping
CC enzymes are restricted to the nucleus (Probable). The enzymatic
CC reaction involves a covalent link between 7-methyl-GMP and nsP1,
CC whereas eukaryotic capping enzymes form a covalent complex only with
CC GMP (By similarity). NsP1 capping consists in the following reactions:
CC GTP is first methylated into 7-methyl-GMP and then is covalently linked
CC to nsP1 to form the m7GMp-nsP1 complex from which 7-methyl-GMP complex
CC is transferred to the mRNA to create the cap structure
CC (PubMed:26041283). NsP1 is also needed for the initiation of the minus-
CC strand RNAs synthesis (By similarity). Probably serves as a membrane
CC anchor for the replication complex composed of nsP1-nsP4 (By
CC similarity). Nsp1 is needed for the initiation of the minus-strand RNAs
CC synthesis (By similarity). Palmitoylated nsP1 is remodeling host cell
CC cytoskeleton, and induces filopodium-like structure formation at the
CC surface of the host cell (By similarity).
CC {ECO:0000250|UniProtKB:P03317, ECO:0000250|UniProtKB:P08411,
CC ECO:0000250|UniProtKB:Q8JUX6, ECO:0000269|PubMed:26041283,
CC ECO:0000305}.
CC -!- FUNCTION: [Protease nsP2]: Multifunctional protein whose N-terminus is
CC part of the RNA polymerase complex and displays NTPase, RNA
CC triphosphatase and helicase activities (By similarity). NTPase and RNA
CC triphosphatase are involved in viral RNA capping and helicase keeps a
CC check on the dsRNA replication intermediates (By similarity). The C-
CC terminus harbors a protease that specifically cleaves the polyproteins
CC and releases the mature proteins (By similarity). Required for the
CC shutoff of minus-strand RNAs synthesis (By similarity). Inhibits host
CC translation to ensure maximal viral gene expression and evade host
CC immune response (PubMed:27318152). {ECO:0000250|UniProtKB:P03317,
CC ECO:0000250|UniProtKB:P08411, ECO:0000250|UniProtKB:Q8JUX6,
CC ECO:0000269|PubMed:27318152}.
CC -!- FUNCTION: [Non-structural protein 3]: Seems to be essential for minus-
CC strand RNAs and subgenomic 26S mRNAs synthesis (By similarity).
CC Displays mono-ADP-ribosylhydrolase activity (PubMed:28150709,
CC PubMed:27440879). ADP-ribosylation is a post-translational modification
CC that controls various processes of the host cell and the virus probably
CC needs to revert it for optimal viral replication (PubMed:28150709).
CC Binds proteins of FXR family and sequesters them into the viral RNA
CC replication complexes thereby inhibiting the formation of host stress
CC granules on viral mRNAs (PubMed:27509095). The nsp3-FXR complexes bind
CC viral RNAs and probably orchestrate the assembly of viral replication
CC complexes, thanks to the ability of FXR family members to self-assemble
CC and bind DNA (PubMed:27509095). {ECO:0000250|UniProtKB:P03317,
CC ECO:0000269|PubMed:27440879, ECO:0000269|PubMed:27509095,
CC ECO:0000269|PubMed:28150709}.
CC -!- FUNCTION: [Non-structural protein 3']: Seems to be essential for minus-
CC strand RNAs and subgenomic 26S mRNAs synthesis (By similarity).
CC Displays mono-ADP-ribosylhydrolase activity (Probable). ADP-
CC ribosylation is a post-translational modification that controls various
CC processes of the host cell and the virus probably needs to revert it
CC for optimal viral replication (Probable). Binds proteins of FXR family
CC and sequesters them into the viral RNA replication complexes thereby
CC inhibiting the formation of host stress granules on viral mRNAs
CC (Probable). The nsp3'-FXR complexes bind viral RNAs and probably
CC orchestrate the assembly of viral replication complexes, thanks to the
CC ability of FXR family members to self-assemble and bind DNA (Probable).
CC {ECO:0000250|UniProtKB:P03317, ECO:0000305|PubMed:27440879,
CC ECO:0000305|PubMed:27509095, ECO:0000305|PubMed:28150709}.
CC -!- FUNCTION: [RNA-directed RNA polymerase nsP4]: RNA dependent RNA
CC polymerase (By similarity). Replicates genomic and antigenomic RNA by
CC recognizing replications specific signals. The early replication
CC complex formed by the polyproteins P123/P123' and nsP4 synthesizes
CC minus-strand RNAs (By similarity). The late replication complex
CC composed of fully processed nsP1-nsP4 is responsible for the production
CC of genomic and subgenomic plus-strand RNAs (By similarity).
CC {ECO:0000250|UniProtKB:P03317}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=GTP + S-adenosyl-L-methionine = N(7)-methyl-GTP + S-adenosyl-
CC L-homocysteine; Xref=Rhea:RHEA:46948, ChEBI:CHEBI:37565,
CC ChEBI:CHEBI:57856, ChEBI:CHEBI:59789, ChEBI:CHEBI:87133;
CC Evidence={ECO:0000269|PubMed:26041283};
CC -!- CATALYTIC ACTIVITY:
CC Reaction=[nsP1 protein]-L-histidine + N(7)-methyl-GTP = [nsP1 protein]-
CC N(tele)-(N(7)-methylguanosine 5'-phospho)-L-histidine + diphosphate;
CC Xref=Rhea:RHEA:54792, Rhea:RHEA-COMP:13994, Rhea:RHEA-COMP:13995,
CC ChEBI:CHEBI:29979, ChEBI:CHEBI:33019, ChEBI:CHEBI:87133,
CC ChEBI:CHEBI:138334; Evidence={ECO:0000269|PubMed:26041283};
CC PhysiologicalDirection=left-to-right; Xref=Rhea:RHEA:54793;
CC Evidence={ECO:0000269|PubMed:26041283};
CC -!- CATALYTIC ACTIVITY:
CC Reaction=[nsP1 protein]-N(tele)-(N(7)-methylguanosine 5'-phospho)-L-
CC histidine + a 5'-end diphospho-(purine-ribonucleoside) in mRNA + H(+)
CC = [nsP1 protein]-L-histidine + a 5'-end (N(7)-methyl 5'-
CC triphosphoguanosine)-(purine-ribonucleoside) in mRNA;
CC Xref=Rhea:RHEA:54800, Rhea:RHEA-COMP:12925, Rhea:RHEA-COMP:13929,
CC Rhea:RHEA-COMP:13994, Rhea:RHEA-COMP:13995, ChEBI:CHEBI:15378,
CC ChEBI:CHEBI:29979, ChEBI:CHEBI:133968, ChEBI:CHEBI:138276,
CC ChEBI:CHEBI:138334; Evidence={ECO:0000269|PubMed:26041283};
CC -!- CATALYTIC ACTIVITY:
CC Reaction=a 5'-end triphospho-(purine-ribonucleoside) in mRNA + H2O = a
CC 5'-end diphospho-(purine-ribonucleoside) in mRNA + H(+) + phosphate;
CC Xref=Rhea:RHEA:11008, Rhea:RHEA-COMP:13929, Rhea:RHEA-COMP:13942,
CC ChEBI:CHEBI:15377, ChEBI:CHEBI:15378, ChEBI:CHEBI:43474,
CC ChEBI:CHEBI:138276, ChEBI:CHEBI:138288; EC=3.1.3.33;
CC Evidence={ECO:0000250|UniProtKB:P08411};
CC -!- CATALYTIC ACTIVITY:
CC Reaction=a ribonucleoside 5'-triphosphate + H2O = a ribonucleoside 5'-
CC diphosphate + H(+) + phosphate; Xref=Rhea:RHEA:23680,
CC ChEBI:CHEBI:15377, ChEBI:CHEBI:15378, ChEBI:CHEBI:43474,
CC ChEBI:CHEBI:57930, ChEBI:CHEBI:61557; EC=3.6.1.15;
CC Evidence={ECO:0000250|UniProtKB:Q8JUX6};
CC -!- CATALYTIC ACTIVITY:
CC Reaction=ATP + H2O = ADP + H(+) + phosphate; Xref=Rhea:RHEA:13065,
CC ChEBI:CHEBI:15377, ChEBI:CHEBI:15378, ChEBI:CHEBI:30616,
CC ChEBI:CHEBI:43474, ChEBI:CHEBI:456216; EC=3.6.4.13;
CC Evidence={ECO:0000250|UniProtKB:Q8JUX6};
CC -!- CATALYTIC ACTIVITY:
CC Reaction=a ribonucleoside 5'-triphosphate + RNA(n) = diphosphate +
CC RNA(n+1); Xref=Rhea:RHEA:21248, Rhea:RHEA-COMP:14527, Rhea:RHEA-
CC COMP:17342, ChEBI:CHEBI:33019, ChEBI:CHEBI:61557, ChEBI:CHEBI:140395;
CC EC=2.7.7.48; Evidence={ECO:0000255|PROSITE-ProRule:PRU00539};
CC -!- CATALYTIC ACTIVITY:
CC Reaction=4-O-(ADP-D-ribosyl)-L-aspartyl-[protein] + H2O = ADP-D-ribose
CC + H(+) + L-aspartyl-[protein]; Xref=Rhea:RHEA:54428, Rhea:RHEA-
CC COMP:9867, Rhea:RHEA-COMP:13832, ChEBI:CHEBI:15377,
CC ChEBI:CHEBI:15378, ChEBI:CHEBI:29961, ChEBI:CHEBI:57967,
CC ChEBI:CHEBI:138102; Evidence={ECO:0000269|PubMed:28150709};
CC PhysiologicalDirection=left-to-right; Xref=Rhea:RHEA:54429;
CC Evidence={ECO:0000269|PubMed:28150709};
CC -!- CATALYTIC ACTIVITY:
CC Reaction=5-O-(ADP-D-ribosyl)-L-glutamyl-[protein] + H2O = ADP-D-ribose
CC + H(+) + L-glutamyl-[protein]; Xref=Rhea:RHEA:58248, Rhea:RHEA-
CC COMP:10208, Rhea:RHEA-COMP:15089, ChEBI:CHEBI:15377,
CC ChEBI:CHEBI:15378, ChEBI:CHEBI:29973, ChEBI:CHEBI:57967,
CC ChEBI:CHEBI:142540; Evidence={ECO:0000269|PubMed:27440879,
CC ECO:0000269|PubMed:28150709};
CC PhysiologicalDirection=left-to-right; Xref=Rhea:RHEA:58249;
CC Evidence={ECO:0000269|PubMed:27440879, ECO:0000269|PubMed:28150709};
CC -!- CATALYTIC ACTIVITY:
CC Reaction=ATP + RNA(n) = diphosphate + RNA(n)-3'-adenine ribonucleotide;
CC Xref=Rhea:RHEA:11332, Rhea:RHEA-COMP:14527, Rhea:RHEA-COMP:17347,
CC ChEBI:CHEBI:30616, ChEBI:CHEBI:33019, ChEBI:CHEBI:140395,
CC ChEBI:CHEBI:173115; EC=2.7.7.19;
CC Evidence={ECO:0000250|UniProtKB:P03317};
CC -!- CATALYTIC ACTIVITY:
CC Reaction=ADP-beta-D-ribose 1''-phosphate + H2O = ADP-D-ribose +
CC phosphate; Xref=Rhea:RHEA:25029, ChEBI:CHEBI:15377,
CC ChEBI:CHEBI:43474, ChEBI:CHEBI:57967, ChEBI:CHEBI:58753; EC=3.1.3.84;
CC Evidence={ECO:0000250|UniProtKB:P36328};
CC PhysiologicalDirection=left-to-right; Xref=Rhea:RHEA:25030;
CC Evidence={ECO:0000250|UniProtKB:P36328};
CC -!- COFACTOR:
CC Name=Mg(2+); Xref=ChEBI:CHEBI:18420;
CC Evidence={ECO:0000250|UniProtKB:P03317};
CC Name=Mn(2+); Xref=ChEBI:CHEBI:29035;
CC Evidence={ECO:0000250|UniProtKB:P03317};
CC Note=For nsP4 adenylyltransferase activity; Mn(2+) supports catalysis
CC at 60% of the levels observed with Mg(2+).
CC {ECO:0000250|UniProtKB:P03317};
CC -!- COFACTOR:
CC Name=Mg(2+); Xref=ChEBI:CHEBI:18420;
CC Evidence={ECO:0000250|UniProtKB:P03317};
CC Note=For nsP4 RNA-directed RNA polymerase activity.
CC {ECO:0000250|UniProtKB:P03317};
CC -!- COFACTOR:
CC Name=Mg(2+); Xref=ChEBI:CHEBI:18420;
CC Evidence={ECO:0000269|PubMed:26041283};
CC Note=For nsP1 guanylylation. {ECO:0000269|PubMed:26041283};
CC -!- COFACTOR:
CC Name=Mg(2+); Xref=ChEBI:CHEBI:18420;
CC Note=For nsP2 RNA triphosphatase activity.
CC {ECO:0000250|UniProtKB:Q8JUX6};
CC -!- COFACTOR:
CC Name=Mg(2+); Xref=ChEBI:CHEBI:18420;
CC Note=For nsP2 NTPase activity. {ECO:0000250|UniProtKB:Q8JUX6};
CC -!- ACTIVITY REGULATION: [mRNA-capping enzyme nsP1]: Inhibited by
CC sinefungin. {ECO:0000269|PubMed:26041283}.
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=110 uM for a 12 residues substrate for nsp2 protease activity
CC {ECO:0000269|PubMed:27030368};
CC pH dependence:
CC Optimum pH is 7 for nsP1 guanylylation.
CC {ECO:0000269|PubMed:26041283};
CC -!- SUBUNIT: [mRNA-capping enzyme nsP1]: Interacts with non-structural
CC protein 3 (By similarity). Interacts with RNA-directed RNA polymerase
CC nsP4 (By similarity). Interacts with protease nsP2 (By similarity).
CC interacts with itself (By similarity). {ECO:0000250|UniProtKB:Q8JUX6}.
CC -!- SUBUNIT: [Non-structural protein 3]: Interacts with mRNA-capping enzyme
CC nsP1 (By similarity). Interacts with host DDX1 (PubMed:27105836).
CC Interacts with host DDX3 (PubMed:27105836). Interacts (via C-terminus)
CC with host FXR1; this interaction inhibits the formation of host stress
CC granules on viral mRNAs and the nsp3-FXR1 complexes bind viral RNAs and
CC probably orchestrate the assembly of viral replication complexes
CC (PubMed:27509095). Interacts (via C-terminus) with host FXR2; this
CC interaction inhibits the formation of host stress granules on viral
CC mRNAs and the nsp3-FXR2 complexes bind viral RNAs and probably
CC orchestrate the assembly of viral replication complexes
CC (PubMed:27509095). Interacts (via C-terminus) with host FMR1; this
CC interaction inhibits the formation of host stress granules on viral
CC mRNAs and the nsp3-FMR1 complexes bind viral RNAs and probably
CC orchestrate the assembly of viral replication complexes
CC (PubMed:27509095). {ECO:0000250|UniProtKB:Q8JUX6,
CC ECO:0000269|PubMed:27105836, ECO:0000269|PubMed:27509095}.
CC -!- SUBUNIT: [RNA-directed RNA polymerase nsP4]: Interacts with mRNA-
CC capping enzyme nsP1 (By similarity). Interacts with protease nsP2 (By
CC similarity). interacts with itself (By similarity).
CC {ECO:0000250|UniProtKB:Q8JUX6}.
CC -!- SUBUNIT: [Protease nsP2]: Interacts with RNA-directed RNA polymerase
CC nsP4 (By similarity). Interacts with mRNA-capping enzyme nsP1 (By
CC similarity). Interacts with KPNA1/karyopherin-alpha1; this interaction
CC probably allows the active transport of protease nsP2 into the host
CC nucleus (PubMed:17652399). {ECO:0000250|UniProtKB:Q8JUX6,
CC ECO:0000269|PubMed:17652399}.
CC -!- SUBCELLULAR LOCATION: [Polyprotein P1234]: Host cytoplasmic vesicle
CC membrane {ECO:0000305}; Peripheral membrane protein {ECO:0000305}.
CC Note=Part of cytoplasmic vesicles, which are probably formed at the
CC plasma membrane and internalized leading to late endosomal/lysosomal
CC spherules containing the replication complex. {ECO:0000305}.
CC -!- SUBCELLULAR LOCATION: [Polyprotein P123']: Host cytoplasmic vesicle
CC membrane {ECO:0000305}; Peripheral membrane protein {ECO:0000305}.
CC Note=Part of cytoplasmic vesicles, which are probably formed at the
CC plasma membrane and internalized leading to late endosomal/lysosomal
CC spherules containing the replication complex. {ECO:0000305}.
CC -!- SUBCELLULAR LOCATION: [Polyprotein P123]: Host cytoplasmic vesicle
CC membrane {ECO:0000305}; Peripheral membrane protein {ECO:0000305}.
CC Note=Part of cytoplasmic vesicles, which are probably formed at the
CC plasma membrane and internalized leading to late endosomal/lysosomal
CC spherules containing the replication complex. {ECO:0000305}.
CC -!- SUBCELLULAR LOCATION: [mRNA-capping enzyme nsP1]: Host cytoplasmic
CC vesicle membrane {ECO:0000250|UniProtKB:P08411}; Lipid-anchor
CC {ECO:0000250|UniProtKB:P08411}. Host cell membrane
CC {ECO:0000250|UniProtKB:P08411}; Lipid-anchor
CC {ECO:0000250|UniProtKB:P08411}; Cytoplasmic side
CC {ECO:0000250|UniProtKB:P08411}. Host cell projection, host filopodium
CC {ECO:0000250|UniProtKB:P08411}. Note=In the late phase of infection,
CC the polyprotein is quickly cleaved before localization to cellular
CC membranes. Then a fraction of nsP1 localizes to the inner surface of
CC the plasma membrane and its filopodial extensions. Only the
CC palmitoylated nsP1 localizes to the host filopodia (By similarity).
CC NsP1 is also part of cytoplasmic vesicles, which are probably formed at
CC the plasma membrane and internalized leading to late
CC endosomal/lysosomal spherules containing the replication complex (By
CC similarity). {ECO:0000250|UniProtKB:P08411}.
CC -!- SUBCELLULAR LOCATION: [Protease nsP2]: Host cytoplasmic vesicle
CC membrane {ECO:0000250|UniProtKB:P08411}; Peripheral membrane protein
CC {ECO:0000250|UniProtKB:P08411}. Host nucleus
CC {ECO:0000269|PubMed:17652399}. Host cytoplasm
CC {ECO:0000269|PubMed:17652399}. Note=In the late phase of infection, the
CC polyprotein is quickly cleaved before localization to cellular
CC membranes. Then approximately half of nsP2 is found in the nucleus (By
CC similarity). Shuttles between cytoplasm and nucleus (PubMed:17652399).
CC NsP2 is also part of cytoplasmic vesicles, which are probably formed at
CC the plasma membrane and internalized leading to late
CC endosomal/lysosomal spherules containing the replication complex (By
CC similarity). {ECO:0000250|UniProtKB:P08411,
CC ECO:0000269|PubMed:17652399}.
CC -!- SUBCELLULAR LOCATION: [Non-structural protein 3]: Host cytoplasmic
CC vesicle membrane {ECO:0000250|UniProtKB:P03317}; Peripheral membrane
CC protein {ECO:0000305}. Note=In the late phase of infection, the
CC polyprotein is quickly cleaved before localization to cellular
CC membranes. Then nsP3 and nsP3' form aggregates in cytoplasm (By
CC similarity). NsP3 is also part of cytoplasmic vesicles, which are
CC probably formed at the plasma membrane and internalized leading to late
CC endosomal/lysosomal spherules containing the replication complex (By
CC similarity). {ECO:0000250|UniProtKB:P03317}.
CC -!- SUBCELLULAR LOCATION: [Non-structural protein 3']: Host cytoplasmic
CC vesicle membrane {ECO:0000305}; Peripheral membrane protein
CC {ECO:0000305}. Note=In the late phase of infection, the polyprotein is
CC quickly cleaved before localization to cellular membranes. Then nsP3
CC and nsP3' form aggregates in cytoplasm (By similarity). NsP3' is also
CC part of cytoplasmic vesicles, which are probably formed at the plasma
CC membrane and internalized leading to late endosomal/lysosomal spherules
CC containing the replication complex (Probable).
CC {ECO:0000250|UniProtKB:P03317, ECO:0000305}.
CC -!- SUBCELLULAR LOCATION: [RNA-directed RNA polymerase nsP4]: Host
CC cytoplasmic vesicle membrane; Peripheral membrane protein
CC {ECO:0000250|UniProtKB:P08411}. Note=NsP4 is part of cytoplasmic
CC vesicles, which are probably formed at the plasma membrane and
CC internalized leading to late endosomal/lysosomal spherules containing
CC the replication complex. {ECO:0000250|UniProtKB:P08411}.
CC -!- DOMAIN: [Protease nsP2]: The N-terminus exhibits NTPase and RNA
CC triphosphatase activities and is proposed to have helicase activity,
CC whereas the C-terminus possesses protease activity (By similarity).
CC Contains a nuclear localization signal and a nuclear export signal,
CC these two motifs are probably involved in the shuttling between the
CC cytoplasm and the nucleus of nsP2 (PubMed:17652399).
CC {ECO:0000250|UniProtKB:Q8JUX6, ECO:0000269|PubMed:17652399}.
CC -!- DOMAIN: [Non-structural protein 3]: In the N-terminus, the macro domain
CC displays a mono-ADP-ribosylhydrolase activity (PubMed:28150709,
CC PubMed:27440879). The central part has a zinc-binding function (By
CC similarity). The C-terminus contains two approximate repeats necessary
CC and sufficient for formation of the nsP3/FXR complex (PubMed:27509095).
CC {ECO:0000250|UniProtKB:P03317, ECO:0000269|PubMed:27440879,
CC ECO:0000269|PubMed:27509095, ECO:0000269|PubMed:28150709}.
CC -!- DOMAIN: [Non-structural protein 3']: In the N-terminus, the macro
CC domain displays a mono-ADP-ribosylhydrolase activity (Probable). The
CC central part has a zinc-binding function (By similarity). The C-
CC terminus contains two approximate repeats necessary and sufficient for
CC formation of the nsP3'/FXR complex (Probable).
CC {ECO:0000250|UniProtKB:P03317, ECO:0000305|PubMed:27440879,
CC ECO:0000305|PubMed:27509095, ECO:0000305|PubMed:28150709}.
CC -!- PTM: [Polyprotein P1234]: Specific enzymatic cleavages in vivo yield
CC mature proteins (By similarity). The processing of the polyprotein is
CC temporally regulated (By similarity). In early stages (1.7 hpi), P1234
CC is first cleaved in trans through its nsP2 protease activity, releasing
CC P123' and nsP4, which associate to form the early replication complex
CC (By similarity). At the same time, P1234 is also cut at the nsP1/nsP2
CC site early in infection but with lower efficiency (By similarity).
CC After replication of the viral minus-strand RNAs (4 hpi), the
CC polyproteins are cut at the nsP1/nsP2 and nsP2/nsP3 sites very
CC efficiently, preventing accumulation of P123' and P1234 and allowing
CC the formation of the late replication complex (By similarity).
CC NsP3'/nsP4 site is not cleaved anymore and P34 is produced rather than
CC nsP4 (By similarity). {ECO:0000250|UniProtKB:P03317}.
CC -!- PTM: [Polyprotein P123]: Specific enzymatic cleavages in vivo yield
CC mature proteins (By similarity). The processing of the polyprotein is
CC temporally regulated (By similarity). In early stages (1.7 hpi), P123
CC is cleaved at the nsP1/nsP2 site with low efficiency (By similarity).
CC After replication of the viral minus-strand RNAs (4 hpi), the
CC polyproteins are cut at the nsP1/nsP2 and nsP2/nsP3 sites very
CC efficiently, preventing accumulation of P123 and allowing the formation
CC of the late replication complex (By similarity).
CC {ECO:0000250|UniProtKB:P03317}.
CC -!- PTM: [Polyprotein P123']: Specific enzymatic cleavages in vivo yield
CC mature proteins (By similarity). The processing of the polyprotein is
CC temporally regulated (By similarity). In early stages (1.7 hpi), P123'
CC is cleaved at the nsP1/nsP2 site with low efficiency (By similarity).
CC After replication of the viral minus-strand RNAs (4 hpi), the
CC polyproteins are cut at the nsP1/nsP2 and nsP2/nsP3 sites very
CC efficiently, preventing accumulation of P123' and allowing the
CC formation of the late replication complex (By similarity).
CC {ECO:0000250|UniProtKB:P03317}.
CC -!- PTM: [mRNA-capping enzyme nsP1]: Palmitoylated by host
CC palmitoyltransferases ZDHHC2 and ZDHHC19.
CC {ECO:0000250|UniProtKB:P03317}.
CC -!- PTM: [Non-structural protein 3]: Phosphorylated by host on serines and
CC threonines. {ECO:0000250|UniProtKB:P08411}.
CC -!- PTM: [Non-structural protein 3']: Phosphorylated by host on serines and
CC threonines. {ECO:0000250|UniProtKB:P08411}.
CC -!- PTM: [RNA-directed RNA polymerase nsP4]: Ubiquitinated; targets the
CC protein for rapid degradation via the ubiquitin system (By similarity).
CC Nsp4 is present in extremely low quantities due to low frequency of
CC translation through the amber stop-codon and the degradation by the
CC ubiquitin pathway (By similarity). {ECO:0000250|UniProtKB:P03317}.
CC -!- MISCELLANEOUS: Viral replication produces dsRNA in the late phase of
CC infection, resulting in a strong activation of host EIF2AK2/PKR,
CC leading to almost complete phosphorylation of EIF2A (By similarity).
CC This inactivates completely cellular translation initiation, resulting
CC shutoff of host proteins synthesis (By similarity). However,
CC phosphorylation of EIF2A is probably not the only mechanism responsible
CC for the host translation shutoff (By similarity). The viral translation
CC can still occur normally because it relies on a hairpin structure in
CC the coding region of sgRNA and is EIF2A-, EIF2D-, EIF4G- EIF4A-
CC independent (By similarity). {ECO:0000250|UniProtKB:P03317}.
CC -!- MISCELLANEOUS: [Polyprotein P1234]: The genome codes for P123, but
CC readthrough of a terminator codon UGA occurs between the codons for
CC Gln-1879 and Arg-1881 giving rise to P1234 (By similarity). P1234 is
CC cleaved quickly by nsP2 into P123' and nsP4 (By similarity). Further
CC processing of p123' gives nsP1, nsP2 and nsP3' which is 6 amino acids
CC longer than nsP3 since the cleavage site is after the readthrough (By
CC similarity). This unusual molecular mechanism ensures that few nsP4 are
CC produced compared to other non-structural proteins (By similarity).
CC Mutant viruses with no alternative termination site grow significantly
CC slower than wild-type virus (By similarity). The opal termination codon
CC is frequently mutated to a sense codon on passage in cell culture (By
CC similarity). The presence of the opal codon may be a requirement for
CC viral maintenance in both vertebrate and invertebrate hosts and a
CC selective advantage may be conferred in cell culture for the sense
CC codon (By similarity). {ECO:0000250|UniProtKB:O90368,
CC ECO:0000250|UniProtKB:P03317}.
CC -!- SEQUENCE CAUTION:
CC Sequence=ALE15112.1; Type=Erroneous initiation; Note=Truncated N-terminus.; Evidence={ECO:0000305};
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DR EMBL; J04332; AAB02518.1; -; Genomic_RNA.
DR EMBL; L01442; AAC19321.2; -; Genomic_RNA.
DR EMBL; L01443; AAB02516.1; -; Genomic_RNA.
DR EMBL; AF069903; AAC24033.1; -; Genomic_RNA.
DR EMBL; AY741139; AAU89533.1; -; Genomic_RNA.
DR EMBL; KR260736; ALE15112.1; ALT_INIT; Genomic_RNA.
DR PIR; A31467; MNWVTD.
DR PDB; 2HWK; X-ray; 2.45 A; A=1003-1322.
DR PDB; 5EZQ; X-ray; 1.66 A; A=992-1327.
DR PDB; 5EZS; X-ray; 2.16 A; A=992-1327.
DR PDB; 6BCM; X-ray; 2.10 A; A=1003-1338.
DR PDBsum; 2HWK; -.
DR PDBsum; 5EZQ; -.
DR PDBsum; 5EZS; -.
DR PDBsum; 6BCM; -.
DR SMR; P27282; -.
DR ChEMBL; CHEMBL4739846; -.
DR MEROPS; C09.002; -.
DR MEROPS; S03.001; -.
DR PRIDE; P27282; -.
DR BRENDA; 3.4.22.B79; 9640.
DR EvolutionaryTrace; P27282; -.
DR Proteomes; UP000008659; Genome.
DR Proteomes; UP000100999; Genome.
DR Proteomes; UP000127220; Genome.
DR Proteomes; UP000131973; Genome.
DR Proteomes; UP000146452; Genome.
DR Proteomes; UP000158519; Genome.
DR GO; GO:0044162; C:host cell cytoplasmic vesicle membrane; IEA:UniProtKB-SubCell.
DR GO; GO:0044176; C:host cell filopodium; IEA:UniProtKB-SubCell.
DR GO; GO:0042025; C:host cell nucleus; IEA:UniProtKB-SubCell.
DR GO; GO:0020002; C:host cell plasma membrane; IEA:UniProtKB-SubCell.
DR GO; GO:0016020; C:membrane; IEA:UniProtKB-KW.
DR GO; GO:0047407; F:ADP-ribosyl-[dinitrogen reductase] hydrolase activity; IDA:UniProtKB.
DR GO; GO:0005524; F:ATP binding; IEA:UniProtKB-KW.
DR GO; GO:0016887; F:ATP hydrolysis activity; IEA:RHEA.
DR GO; GO:0008234; F:cysteine-type peptidase activity; IEA:UniProtKB-KW.
DR GO; GO:0005525; F:GTP binding; IEA:UniProtKB-KW.
DR GO; GO:0046872; F:metal ion binding; IEA:UniProtKB-KW.
DR GO; GO:0008174; F:mRNA methyltransferase activity; IEA:InterPro.
DR GO; GO:0004651; F:polynucleotide 5'-phosphatase activity; IEA:UniProtKB-EC.
DR GO; GO:0004652; F:polynucleotide adenylyltransferase activity; IEA:UniProtKB-EC.
DR GO; GO:0003723; F:RNA binding; IEA:UniProtKB-KW.
DR GO; GO:0003724; F:RNA helicase activity; IEA:UniProtKB-EC.
DR GO; GO:0003968; F:RNA-directed 5'-3' RNA polymerase activity; IEA:UniProtKB-KW.
DR GO; GO:0006370; P:7-methylguanosine mRNA capping; IDA:UniProtKB.
DR GO; GO:0006508; P:proteolysis; IEA:UniProtKB-KW.
DR GO; GO:0039523; P:suppression by virus of host mRNA transcription via inhibition of RNA polymerase II activity; IEA:UniProtKB-KW.
DR GO; GO:0006351; P:transcription, DNA-templated; IEA:InterPro.
DR GO; GO:0039694; P:viral RNA genome replication; IEA:InterPro.
DR CDD; cd21557; Macro_X_Nsp3-like; 1.
DR Gene3D; 3.40.220.10; -; 1.
DR Gene3D; 3.40.50.150; -; 1.
DR Gene3D; 3.40.50.300; -; 2.
DR Gene3D; 3.90.70.110; -; 1.
DR InterPro; IPR027351; (+)RNA_virus_helicase_core_dom.
DR InterPro; IPR002588; Alphavirus-like_MT_dom.
DR InterPro; IPR002620; Alphavirus_nsp2pro.
DR InterPro; IPR044936; Alphavirus_nsp2pro_sf.
DR InterPro; IPR043502; DNA/RNA_pol_sf.
DR InterPro; IPR002589; Macro_dom.
DR InterPro; IPR043472; Macro_dom-like.
DR InterPro; IPR044371; Macro_X_NSP3-like.
DR InterPro; IPR027417; P-loop_NTPase.
DR InterPro; IPR007094; RNA-dir_pol_PSvirus.
DR InterPro; IPR029063; SAM-dependent_MTases_sf.
DR InterPro; IPR001788; Tymovirus_RNA-dep_RNA_pol.
DR Pfam; PF01661; Macro; 1.
DR Pfam; PF01707; Peptidase_C9; 1.
DR Pfam; PF00978; RdRP_2; 1.
DR Pfam; PF01443; Viral_helicase1; 1.
DR Pfam; PF01660; Vmethyltransf; 1.
DR SMART; SM00506; A1pp; 1.
DR SUPFAM; SSF52540; SSF52540; 1.
DR SUPFAM; SSF52949; SSF52949; 1.
DR SUPFAM; SSF56672; SSF56672; 1.
DR PROSITE; PS51743; ALPHAVIRUS_MT; 1.
DR PROSITE; PS51154; MACRO; 1.
DR PROSITE; PS51520; NSP2PRO; 1.
DR PROSITE; PS51657; PSRV_HELICASE; 1.
DR PROSITE; PS50507; RDRP_SSRNA_POS; 1.
PE 1: Evidence at protein level;
KW 3D-structure; ATP-binding;
KW Eukaryotic host gene expression shutoff by virus;
KW Eukaryotic host transcription shutoff by virus; GTP-binding; Helicase;
KW Host cell membrane; Host cell projection; Host cytoplasm;
KW Host cytoplasmic vesicle; Host gene expression shutoff by virus;
KW Host membrane; Host nucleus; Host-virus interaction; Hydrolase;
KW Inhibition of host RNA polymerase II by virus; Lipoprotein; Membrane;
KW Metal-binding; Methyltransferase; mRNA capping; mRNA processing;
KW Multifunctional enzyme; Nucleotide-binding; Nucleotidyltransferase;
KW Palmitate; Phosphoprotein; Protease; Repeat;
KW RNA suppression of termination; RNA-binding; RNA-directed RNA polymerase;
KW S-adenosyl-L-methionine; Thiol protease; Transferase; Ubl conjugation;
KW Viral RNA replication; Zinc.
FT CHAIN 1..2493
FT /note="Polyprotein P1234"
FT /id="PRO_0000308391"
FT CHAIN 1..1886
FT /note="Polyprotein P123'"
FT /id="PRO_0000228770"
FT CHAIN 1..1879
FT /note="Polyprotein P123"
FT /id="PRO_0000228771"
FT CHAIN 1..535
FT /note="mRNA-capping enzyme nsP1"
FT /id="PRO_0000041208"
FT CHAIN 536..1329
FT /note="Protease nsP2"
FT /id="PRO_0000041209"
FT CHAIN 1330..1886
FT /note="Non-structural protein 3'"
FT /id="PRO_0000228772"
FT CHAIN 1330..1879
FT /note="Non-structural protein 3"
FT /id="PRO_0000041210"
FT CHAIN 1887..2493
FT /note="RNA-directed RNA polymerase nsP4"
FT /id="PRO_0000041211"
FT DOMAIN 28..259
FT /note="Alphavirus-like MT"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU01079"
FT DOMAIN 690..841
FT /note="(+)RNA virus helicase ATP-binding"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU00990"
FT DOMAIN 842..990
FT /note="(+)RNA virus helicase C-terminal"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU00990"
FT DOMAIN 1003..1322
FT /note="Peptidase C9"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU00853,
FT ECO:0000269|PubMed:27318152"
FT DOMAIN 1330..1489
FT /note="Macro"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU00490,
FT ECO:0000269|PubMed:28150709"
FT REPEAT 1818..1839
FT /note="1"
FT /evidence="ECO:0000305|PubMed:27509095"
FT REPEAT 1852..1873
FT /note="2"
FT /evidence="ECO:0000305|PubMed:27509095"
FT DOMAIN 2250..2365
FT /note="RdRp catalytic"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU00539"
FT REGION 244..263
FT /note="NsP1 membrane-binding"
FT /evidence="ECO:0000250|UniProtKB:P08411"
FT REGION 1004..1023
FT /note="Nucleolus localization signal"
FT /evidence="ECO:0000250|UniProtKB:P08411"
FT REGION 1660..1696
FT /note="Disordered"
FT /evidence="ECO:0000256|SAM:MobiDB-lite"
FT REGION 1791..1812
FT /note="Disordered"
FT /evidence="ECO:0000256|SAM:MobiDB-lite"
FT REGION 1818..1873
FT /note="2 X 21 AA approximate repeats, binding to host FXR
FT family members"
FT /evidence="ECO:0000269|PubMed:27509095"
FT MOTIF 1056..1065
FT /note="Nuclear export signal"
FT /evidence="ECO:0000269|PubMed:17652399"
FT MOTIF 1179..1183
FT /note="Nuclear localization signal"
FT /evidence="ECO:0000269|PubMed:17652399"
FT COMPBIAS 1669..1684
FT /note="Polar residues"
FT /evidence="ECO:0000256|SAM:MobiDB-lite"
FT COMPBIAS 1792..1806
FT /note="Pro residues"
FT /evidence="ECO:0000256|SAM:MobiDB-lite"
FT ACT_SITE 1012
FT /note="For cysteine protease nsP2 activity"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU00853,
FT ECO:0000269|PubMed:16962975"
FT ACT_SITE 1081
FT /note="For cysteine protease nsP2 activity"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU00853,
FT ECO:0000269|PubMed:16962975"
FT BINDING 721..728
FT /ligand="a ribonucleoside 5'-triphosphate"
FT /ligand_id="ChEBI:CHEBI:61557"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU00990"
FT BINDING 1339
FT /ligand="ADP-D-ribose"
FT /ligand_id="ChEBI:CHEBI:57967"
FT /evidence="ECO:0000250|UniProtKB:P36328"
FT BINDING 1353
FT /ligand="ADP-D-ribose"
FT /ligand_id="ChEBI:CHEBI:57967"
FT /evidence="ECO:0000250|UniProtKB:Q8JUX6"
FT BINDING 1361
FT /ligand="ADP-D-ribose"
FT /ligand_id="ChEBI:CHEBI:57967"
FT /evidence="ECO:0000250|UniProtKB:Q8JUX6"
FT BINDING 1441
FT /ligand="ADP-D-ribose"
FT /ligand_id="ChEBI:CHEBI:57967"
FT /evidence="ECO:0000250|UniProtKB:P36328"
FT BINDING 1442
FT /ligand="ADP-D-ribose"
FT /ligand_id="ChEBI:CHEBI:57967"
FT /evidence="ECO:0000250|UniProtKB:P36328"
FT BINDING 1443
FT /ligand="ADP-D-ribose"
FT /ligand_id="ChEBI:CHEBI:57967"
FT /evidence="ECO:0000250|UniProtKB:P36328"
FT BINDING 1596
FT /ligand="Zn(2+)"
FT /ligand_id="ChEBI:CHEBI:29105"
FT /evidence="ECO:0000250|UniProtKB:P03317"
FT BINDING 1598
FT /ligand="Zn(2+)"
FT /ligand_id="ChEBI:CHEBI:29105"
FT /evidence="ECO:0000250|UniProtKB:P03317"
FT BINDING 1621
FT /ligand="Zn(2+)"
FT /ligand_id="ChEBI:CHEBI:29105"
FT /evidence="ECO:0000250|UniProtKB:P03317"
FT BINDING 1639
FT /ligand="Zn(2+)"
FT /ligand_id="ChEBI:CHEBI:29105"
FT /evidence="ECO:0000250|UniProtKB:P03317"
FT SITE 37
FT /note="Involved in the phosphoramide link with 7-methyl-
FT GMP"
FT /evidence="ECO:0000269|PubMed:26041283"
FT SITE 535..536
FT /note="Cleavage; by protease nsP2"
FT /evidence="ECO:0000250|UniProtKB:P03317"
FT SITE 1329..1330
FT /note="Cleavage; by protease nsP2"
FT /evidence="ECO:0000250|UniProtKB:P03317"
FT SITE 1886..1887
FT /note="Cleavage; by protease nsP2"
FT /evidence="ECO:0000250|UniProtKB:Q8JUX6"
FT LIPID 419
FT /note="S-palmitoyl cysteine; by host"
FT /evidence="ECO:0000250|UniProtKB:Q8JUX6"
FT VARIANT 222
FT /note="R -> C (in strain: 71-180 and COAN5506)"
FT VARIANT 551
FT /note="A -> D (in strain: TC-83)"
FT MUTAGEN 37
FT /note="H->A: Complete loss of nsP1 guanylylation; increased
FT methyltransferase activity."
FT /evidence="ECO:0000269|PubMed:26041283"
FT MUTAGEN 45
FT /note="H->A: Almost no effect on nsP1 guanylylation; 50%
FT loss of methyltransferase activity."
FT /evidence="ECO:0000269|PubMed:26041283"
FT MUTAGEN 63
FT /note="D->A: Complete loss of nsP1 guanylylation; 80% loss
FT of methyltransferase activity."
FT /evidence="ECO:0000269|PubMed:26041283"
FT MUTAGEN 118
FT /note="E->A: Almost no effect on formation of nsP1
FT guanylylation."
FT /evidence="ECO:0000269|PubMed:26041283"
FT MUTAGEN 285
FT /note="Y->A: 85% loss of formation of nsP1 guanylylation;
FT 70% loss of methyltransferase activity."
FT /evidence="ECO:0000269|PubMed:26041283"
FT MUTAGEN 354
FT /note="D->A: Increased formation of nsP1 guanylylation;
FT slightly increased methyltransferase activity."
FT /evidence="ECO:0000269|PubMed:26041283"
FT MUTAGEN 365
FT /note="R->A: Increased formation of nsP1 guanylylation;
FT slightly increased methyltransferase activity."
FT /evidence="ECO:0000269|PubMed:26041283"
FT MUTAGEN 369
FT /note="N->A: 50% loss of formation of nsP1 guanylylation;
FT 50% loss of methyltransferase activity."
FT /evidence="ECO:0000269|PubMed:26041283"
FT MUTAGEN 375
FT /note="N->A: 50% loss of formation of nsP1 guanylylation;
FT 40% loss of methyltransferase activity."
FT /evidence="ECO:0000269|PubMed:26041283"
FT MUTAGEN 538
FT /note="V->A: Increased viral replication and
FT cytopathogenicity."
FT /evidence="ECO:0000269|PubMed:23365438"
FT MUTAGEN 540
FT /note="T->A: Increased viral replication and
FT cytopathogenicity."
FT /evidence="ECO:0000269|PubMed:23365438"
FT MUTAGEN 1006
FT /note="Q->A: Increased viral replication and
FT cytopathogenicity."
FT /evidence="ECO:0000269|PubMed:23365438"
FT MUTAGEN 1010
FT /note="N->A: 24 fold reduced kcat/KM for nsP2 protease
FT activity."
FT /evidence="ECO:0000269|PubMed:27030368"
FT MUTAGEN 1010
FT /note="N->A: NsP2 is in a self-inhibited state."
FT /evidence="ECO:0000269|PubMed:29064679"
FT MUTAGEN 1015
FT /note="K->A: 9 fold reduced kcat/KM for nsP2 protease
FT activity."
FT /evidence="ECO:0000269|PubMed:27030368"
FT MUTAGEN 1056
FT /note="V->A: No effect on nuclear export of nsP2."
FT /evidence="ECO:0000269|PubMed:17652399"
FT MUTAGEN 1061
FT /note="L->A: Inhibits the nuclear export of nsP2; when
FT associated with Ala-1063."
FT /evidence="ECO:0000269|PubMed:17652399"
FT MUTAGEN 1063
FT /note="L->A: Inhibits the nuclear export of nsP2; when
FT associated with Ala-1061."
FT /evidence="ECO:0000269|PubMed:17652399"
FT MUTAGEN 1181
FT /note="K->D: Decreased protease nsP2 nuclear localization."
FT /evidence="ECO:0000269|PubMed:17652399"
FT MUTAGEN 1197
FT /note="R->A,K: 16 fold reduced kcat/KM for nsP2 protease
FT activity."
FT /evidence="ECO:0000269|PubMed:27030368"
FT MUTAGEN 1248
FT /note="P->S: No effect on host shutoff induction by nsP2 at
FT 6h p.i."
FT /evidence="ECO:0000269|PubMed:27318152"
FT MUTAGEN 1274
FT /note="Q->L: 50% loss of host shutoff induction by nsP2 at
FT 6h p.i."
FT /evidence="ECO:0000269|PubMed:27318152"
FT CONFLICT 21
FT /note="S -> T (in Ref. 1; AAB02518)"
FT /evidence="ECO:0000305"
FT CONFLICT 497
FT /note="A -> R (in Ref. 1; AAB02518)"
FT /evidence="ECO:0000305"
FT CONFLICT 1589
FT /note="T -> S (in Ref. 1; AAB02518)"
FT /evidence="ECO:0000305"
FT TURN 1004..1007
FT /evidence="ECO:0007829|PDB:5EZS"
FT HELIX 1013..1022
FT /evidence="ECO:0007829|PDB:6BCM"
FT TURN 1023..1025
FT /evidence="ECO:0007829|PDB:6BCM"
FT HELIX 1030..1033
FT /evidence="ECO:0007829|PDB:6BCM"
FT TURN 1037..1042
FT /evidence="ECO:0007829|PDB:6BCM"
FT HELIX 1047..1059
FT /evidence="ECO:0007829|PDB:6BCM"
FT HELIX 1063..1065
FT /evidence="ECO:0007829|PDB:6BCM"
FT STRAND 1069..1071
FT /evidence="ECO:0007829|PDB:6BCM"
FT STRAND 1076..1078
FT /evidence="ECO:0007829|PDB:6BCM"
FT STRAND 1081..1083
FT /evidence="ECO:0007829|PDB:6BCM"
FT STRAND 1090..1092
FT /evidence="ECO:0007829|PDB:6BCM"
FT HELIX 1095..1101
FT /evidence="ECO:0007829|PDB:6BCM"
FT TURN 1102..1104
FT /evidence="ECO:0007829|PDB:6BCM"
FT HELIX 1108..1114
FT /evidence="ECO:0007829|PDB:6BCM"
FT TURN 1120..1122
FT /evidence="ECO:0007829|PDB:6BCM"
FT HELIX 1158..1162
FT /evidence="ECO:0007829|PDB:6BCM"
FT STRAND 1167..1174
FT /evidence="ECO:0007829|PDB:6BCM"
FT STRAND 1181..1189
FT /evidence="ECO:0007829|PDB:6BCM"
FT STRAND 1192..1195
FT /evidence="ECO:0007829|PDB:6BCM"
FT HELIX 1198..1200
FT /evidence="ECO:0007829|PDB:5EZS"
FT STRAND 1209..1215
FT /evidence="ECO:0007829|PDB:6BCM"
FT HELIX 1224..1237
FT /evidence="ECO:0007829|PDB:6BCM"
FT HELIX 1239..1242
FT /evidence="ECO:0007829|PDB:6BCM"
FT HELIX 1243..1245
FT /evidence="ECO:0007829|PDB:6BCM"
FT STRAND 1246..1256
FT /evidence="ECO:0007829|PDB:6BCM"
FT HELIX 1262..1272
FT /evidence="ECO:0007829|PDB:6BCM"
FT STRAND 1275..1281
FT /evidence="ECO:0007829|PDB:6BCM"
FT STRAND 1292..1299
FT /evidence="ECO:0007829|PDB:6BCM"
FT HELIX 1309..1319
FT /evidence="ECO:0007829|PDB:6BCM"
SQ SEQUENCE 2493 AA; 277887 MW; C9FDF4AB76E242DC CRC64;
MEKVHVDIEE DSPFLRALQR SFPQFEVEAK QVTDNDHANA RAFSHLASKL IETEVDPSDT
ILDIGSAPAR RMYSKHKYHC ICPMRCAEDP DRLYKYATKL KKNCKEITDK ELDKKMKELA
AVMSDPDLET ETMCLHDDES CRYEGQVAVY QDVYAVDGPT SLYHQANKGV RVAYWIGFDT
TPFMFKNLAG AYPSYSTNWA DETVLTARNI GLCSSDVMER SRRGMSILRK KYLKPSNNVL
FSVGSTIYHE KRDLLRSWHL PSVFHLRGKQ NYTCRCETIV SCDGYVVKRI AISPGLYGKP
SGYAATMHRE GFLCCKVTDT LNGERVSFPV CTYVPATLCD QMTGILATDV SADDAQKLLV
GLNQRIVVNG RTQRNTNTMK NYLLPVVAQA FARWAKEYKE DQEDERPLGL RDRQLVMGCC
WAFRRHKITS IYKRPDTQTI IKVNSDFHSF VLPRIGSNTL EIGLRTRIRK MLEEHKEPSP
LITAEDVQEA KCAADEAKEV REAEELRAAL PPLAADVEEP TLEADVDLML QEAGAGSVET
PRGLIKVTSY AGEDKIGSYA VLSPQAVLKS EKLSCIHPLA EQVIVITHSG RKGRYAVEPY
HGKVVVPEGH AIPVQDFQAL SESATIVYNE REFVNRYLHH IATHGGALNT DEEYYKTVKP
SEHDGEYLYD IDRKQCVKKE LVTGLGLTGE LVDPPFHEFA YESLRTRPAA PYQVPTIGVY
GVPGSGKSGI IKSAVTKKDL VVSAKKENCA EIIRDVKKMK GLDVNARTVD SVLLNGCKHP
VETLYIDEAF ACHAGTLRAL IAIIRPKKAV LCGDPKQCGF FNMMCLKVHF NHEICTQVFH
KSISRRCTKS VTSVVSTLFY DKKMRTTNPK ETKIVIDTTG STKPKQDDLI LTCFRGWVKQ
LQIDYKGNEI MTAAASQGLT RKGVYAVRYK VNENPLYAPT SEHVNVLLTR TEDRIVWKTL
AGDPWIKTLT AKYPGNFTAT IEEWQAEHDA IMRHILERPD PTDVFQNKAN VCWAKALVPV
LKTAGIDMTT EQWNTVDYFE TDKAHSAEIV LNQLCVRFFG LDLDSGLFSA PTVPLSIRNN
HWDNSPSPNM YGLNKEVVRQ LSRRYPQLPR AVATGRVYDM NTGTLRNYDP RINLVPVNRR
LPHALVLHHN EHPQSDFSSF VSKLKGRTVL VVGEKLSVPG KMVDWLSDRP EATFRARLDL
GIPGDVPKYD IIFVNVRTPY KYHHYQQCED HAIKLSMLTK KACLHLNPGG TCVSIGYGYA
DRASESIIGA IARQFKFSRV CKPKSSLEET EVLFVFIGYD RKARTHNPYK LSSTLTNIYT
GSRLHEAGCA PSYHVVRGDI ATATEGVIIN AANSKGQPGG GVCGALYKKF PESFDLQPIE
VGKARLVKGA AKHIIHAVGP NFNKVSEVEG DKQLAEAYES IAKIVNDNNY KSVAIPLLST
GIFSGNKDRL TQSLNHLLTA LDTTDADVAI YCRDKKWEMT LKEAVARREA VEEICISDDS
SVTEPDAELV RVHPKSSLAG RKGYSTSDGK TFSYLEGTKF HQAAKDIAEI NAMWPVATEA
NEQVCMYILG ESMSSIRSKC PVEESEASTP PSTLPCLCIH AMTPERVQRL KASRPEQITV
CSSFPLPKYR ITGVQKIQCS QPILFSPKVP AYIHPRKYLV ETPPVDETPE PSAENQSTEG
TPEQPPLITE DETRTRTPEP IIIEEEEEDS ISLLSDGPTH QVLQVEADIH GPPSVSSSSW
SIPHASDFDV DSLSILDTLE GASVTSGATS AETNSYFAKS MEFLARPVPA PRTVFRNPPH
PAPRTRTPSL APSRACSRTS LVSTPPGVNR VITREELEAL TPSRTPSRSV SRTSLVSNPP
GVNRVITREE FEAFVAQQQX RFDAGAYIFS SDTGQGHLQQ KSVRQTVLSE VVLERTELEI
SYAPRLDQEK EELLRKKLQL NPTPANRSRY QSRKVENMKA ITARRILQGL GHYLKAEGKV
ECYRTLHPVP LYSSSVNRAF SSPKVAVEAC NAMLKENFPT VASYCIIPEY DAYLDMVDGA
SCCLDTASFC PAKLRSFPKK HSYLEPTIRS AVPSAIQNTL QNVLAAATKR NCNVTQMREL
PVLDSAAFNV ECFKKYACNN EYWETFKENP IRLTEENVVN YITKLKGPKA AALFAKTHNL
NMLQDIPMDR FVMDLKRDVK VTPGTKHTEE RPKVQVIQAA DPLATAYLCG IHRELVRRLN
AVLLPNIHTL FDMSAEDFDA IIAEHFQPGD CVLETDIASF DKSEDDAMAL TALMILEDLG
VDAELLTLIE AAFGEISSIH LPTKTKFKFG AMMKSGMFLT LFVNTVINIV IASRVLRERL
TGSPCAAFIG DDNIVKGVKS DKLMADRCAT WLNMEVKIID AVVGEKAPYF CGGFILCDSV
TGTACRVADP LKRLFKLGKP LAADDEHDDD RRRALHEEST RWNRVGILSE LCKAVESRYE
TVGTSIIVMA MTTLASSVKS FSYLRGAPIT LYG
