| IED ID | IndEnz0002001482 |
| Enzyme Type ID | protease001482 |
| Protein Name |
Polyprotein P1234 P1234 Non-structural polyprotein Cleaved into: Polyprotein P123' P123' ; Polyprotein P123 P123 ; mRNA-capping enzyme nsP1 EC 2.1.1.- EC 2.7.7.- Non-structural protein 1 ; Protease nsP2 EC 3.1.3.33 EC 3.4.22.- EC 3.6.1.15 EC 3.6.4.13 Non-structural protein 2 nsP2 ; Non-structural protein 3' nsP3' EC 3.1.3.84 ; Non-structural protein 3 nsP3 EC 3.1.3.84 ; RNA-directed RNA polymerase nsP4 EC 2.7.7.19 EC 2.7.7.48 Non-structural protein 4 nsP4 |
| Gene Name | |
| Organism | Venezuelan equine encephalitis virus (strain Trinidad donkey) (VEEV) |
| Taxonomic Lineage | Viruses Riboviria Orthornavirae Kitrinoviricota Alsuviricetes Martellivirales Togaviridae Alphavirus (arboviruses group A) Venezuelan equine encephalitis virus Venezuelan equine encephalitis virus (strain Trinidad donkey) (VEEV) |
| Enzyme Sequence | MEKVHVDIEEDSPFLRALQRSFPQFEVEAKQVTDNDHANARAFSHLASKLIETEVDPSDTILDIGSAPARRMYSKHKYHCICPMRCAEDPDRLYKYATKLKKNCKEITDKELDKKMKELAAVMSDPDLETETMCLHDDESCRYEGQVAVYQDVYAVDGPTSLYHQANKGVRVAYWIGFDTTPFMFKNLAGAYPSYSTNWADETVLTARNIGLCSSDVMERSRRGMSILRKKYLKPSNNVLFSVGSTIYHEKRDLLRSWHLPSVFHLRGKQNYTCRCETIVSCDGYVVKRIAISPGLYGKPSGYAATMHREGFLCCKVTDTLNGERVSFPVCTYVPATLCDQMTGILATDVSADDAQKLLVGLNQRIVVNGRTQRNTNTMKNYLLPVVAQAFARWAKEYKEDQEDERPLGLRDRQLVMGCCWAFRRHKITSIYKRPDTQTIIKVNSDFHSFVLPRIGSNTLEIGLRTRIRKMLEEHKEPSPLITAEDVQEAKCAADEAKEVREAEELRAALPPLAADVEEPTLEADVDLMLQEAGAGSVETPRGLIKVTSYAGEDKIGSYAVLSPQAVLKSEKLSCIHPLAEQVIVITHSGRKGRYAVEPYHGKVVVPEGHAIPVQDFQALSESATIVYNEREFVNRYLHHIATHGGALNTDEEYYKTVKPSEHDGEYLYDIDRKQCVKKELVTGLGLTGELVDPPFHEFAYESLRTRPAAPYQVPTIGVYGVPGSGKSGIIKSAVTKKDLVVSAKKENCAEIIRDVKKMKGLDVNARTVDSVLLNGCKHPVETLYIDEAFACHAGTLRALIAIIRPKKAVLCGDPKQCGFFNMMCLKVHFNHEICTQVFHKSISRRCTKSVTSVVSTLFYDKKMRTTNPKETKIVIDTTGSTKPKQDDLILTCFRGWVKQLQIDYKGNEIMTAAASQGLTRKGVYAVRYKVNENPLYAPTSEHVNVLLTRTEDRIVWKTLAGDPWIKTLTAKYPGNFTATIEEWQAEHDAIMRHILERPDPTDVFQNKANVCWAKALVPVLKTAGIDMTTEQWNTVDYFETDKAHSAEIVLNQLCVRFFGLDLDSGLFSAPTVPLSIRNNHWDNSPSPNMYGLNKEVVRQLSRRYPQLPRAVATGRVYDMNTGTLRNYDPRINLVPVNRRLPHALVLHHNEHPQSDFSSFVSKLKGRTVLVVGEKLSVPGKMVDWLSDRPEATFRARLDLGIPGDVPKYDIIFVNVRTPYKYHHYQQCEDHAIKLSMLTKKACLHLNPGGTCVSIGYGYADRASESIIGAIARQFKFSRVCKPKSSLEETEVLFVFIGYDRKARTHNPYKLSSTLTNIYTGSRLHEAGCAPSYHVVRGDIATATEGVIINAANSKGQPGGGVCGALYKKFPESFDLQPIEVGKARLVKGAAKHIIHAVGPNFNKVSEVEGDKQLAEAYESIAKIVNDNNYKSVAIPLLSTGIFSGNKDRLTQSLNHLLTALDTTDADVAIYCRDKKWEMTLKEAVARREAVEEICISDDSSVTEPDAELVRVHPKSSLAGRKGYSTSDGKTFSYLEGTKFHQAAKDIAEINAMWPVATEANEQVCMYILGESMSSIRSKCPVEESEASTPPSTLPCLCIHAMTPERVQRLKASRPEQITVCSSFPLPKYRITGVQKIQCSQPILFSPKVPAYIHPRKYLVETPPVDETPEPSAENQSTEGTPEQPPLITEDETRTRTPEPIIIEEEEEDSISLLSDGPTHQVLQVEADIHGPPSVSSSSWSIPHASDFDVDSLSILDTLEGASVTSGATSAETNSYFAKSMEFLARPVPAPRTVFRNPPHPAPRTRTPSLAPSRACSRTSLVSTPPGVNRVITREELEALTPSRTPSRSVSRTSLVSNPPGVNRVITREEFEAFVAQQQXRFDAGAYIFSSDTGQGHLQQKSVRQTVLSEVVLERTELEISYAPRLDQEKEELLRKKLQLNPTPANRSRYQSRKVENMKAITARRILQGLGHYLKAEGKVECYRTLHPVPLYSSSVNRAFSSPKVAVEACNAMLKENFPTVASYCIIPEYDAYLDMVDGASCCLDTASFCPAKLRSFPKKHSYLEPTIRSAVPSAIQNTLQNVLAAATKRNCNVTQMRELPVLDSAAFNVECFKKYACNNEYWETFKENPIRLTEENVVNYITKLKGPKAAALFAKTHNLNMLQDIPMDRFVMDLKRDVKVTPGTKHTEERPKVQVIQAADPLATAYLCGIHRELVRRLNAVLLPNIHTLFDMSAEDFDAIIAEHFQPGDCVLETDIASFDKSEDDAMALTALMILEDLGVDAELLTLIEAAFGEISSIHLPTKTKFKFGAMMKSGMFLTLFVNTVINIVIASRVLRERLTGSPCAAFIGDDNIVKGVKSDKLMADRCATWLNMEVKIIDAVVGEKAPYFCGGFILCDSVTGTACRVADPLKRLFKLGKPLAADDEHDDDRRRALHEESTRWNRVGILSELCKAVESRYETVGTSIIVMAMTTLASSVKSFSYLRGAPITLYG |
| Enzyme Length | 2493 |
| Uniprot Accession Number | P27282 |
| Absorption | |
| Active Site | ACT_SITE 1012; /note="For cysteine protease nsP2 activity"; /evidence="ECO:0000255|PROSITE-ProRule:PRU00853, ECO:0000269|PubMed:16962975"; ACT_SITE 1081; /note="For cysteine protease nsP2 activity"; /evidence="ECO:0000255|PROSITE-ProRule:PRU00853, ECO:0000269|PubMed:16962975" |
| Activity Regulation | ACTIVITY REGULATION: [mRNA-capping enzyme nsP1]: Inhibited by sinefungin. {ECO:0000269|PubMed:26041283}. |
| Binding Site | BINDING 1339; /note=ADP-ribose; /evidence=ECO:0000250|UniProtKB:P36328; BINDING 1353; /note=ADP-ribose; /evidence=ECO:0000250|UniProtKB:Q8JUX6; BINDING 1361; /note=ADP-ribose; /evidence=ECO:0000250|UniProtKB:Q8JUX6; BINDING 1441; /note=ADP-ribose; /evidence=ECO:0000250|UniProtKB:P36328; BINDING 1442; /note=ADP-ribose; /evidence=ECO:0000250|UniProtKB:P36328; BINDING 1443; /note=ADP-ribose; /evidence=ECO:0000250|UniProtKB:P36328 |
| Calcium Binding | |
| catalytic Activity | CATALYTIC ACTIVITY: Reaction=GTP + S-adenosyl-L-methionine = N(7)-methyl-GTP + S-adenosyl-L-homocysteine; Xref=Rhea:RHEA:46948, ChEBI:CHEBI:37565, ChEBI:CHEBI:57856, ChEBI:CHEBI:59789, ChEBI:CHEBI:87133; Evidence={ECO:0000269|PubMed:26041283}; CATALYTIC ACTIVITY: Reaction=[nsP1 protein]-L-histidine + N(7)-methyl-GTP = [nsP1 protein]-N(tele)-(N(7)-methylguanosine 5'-phospho)-L-histidine + diphosphate; Xref=Rhea:RHEA:54792, Rhea:RHEA-COMP:13994, Rhea:RHEA-COMP:13995, ChEBI:CHEBI:29979, ChEBI:CHEBI:33019, ChEBI:CHEBI:87133, ChEBI:CHEBI:138334; Evidence={ECO:0000269|PubMed:26041283};PhysiologicalDirection=left-to-right; Xref=Rhea:RHEA:54793; Evidence={ECO:0000269|PubMed:26041283}; CATALYTIC ACTIVITY: Reaction=[nsP1 protein]-N(tele)-(N(7)-methylguanosine 5'-phospho)-L-histidine + a 5'-end diphospho-(purine-ribonucleoside) in mRNA + H(+) = [nsP1 protein]-L-histidine + a 5'-end (N(7)-methyl 5'-triphosphoguanosine)-(purine-ribonucleoside) in mRNA; Xref=Rhea:RHEA:54800, Rhea:RHEA-COMP:12925, Rhea:RHEA-COMP:13929, Rhea:RHEA-COMP:13994, Rhea:RHEA-COMP:13995, ChEBI:CHEBI:15378, ChEBI:CHEBI:29979, ChEBI:CHEBI:133968, ChEBI:CHEBI:138276, ChEBI:CHEBI:138334; Evidence={ECO:0000269|PubMed:26041283}; CATALYTIC ACTIVITY: Reaction=a 5'-end triphospho-(purine-ribonucleoside) in mRNA + H2O = a 5'-end diphospho-(purine-ribonucleoside) in mRNA + H(+) + phosphate; Xref=Rhea:RHEA:11008, Rhea:RHEA-COMP:13929, Rhea:RHEA-COMP:13942, ChEBI:CHEBI:15377, ChEBI:CHEBI:15378, ChEBI:CHEBI:43474, ChEBI:CHEBI:138276, ChEBI:CHEBI:138288; EC=3.1.3.33; Evidence={ECO:0000250|UniProtKB:P08411}; CATALYTIC ACTIVITY: Reaction=a ribonucleoside 5'-triphosphate + H2O = a ribonucleoside 5'-diphosphate + H(+) + phosphate; Xref=Rhea:RHEA:23680, ChEBI:CHEBI:15377, ChEBI:CHEBI:15378, ChEBI:CHEBI:43474, ChEBI:CHEBI:57930, ChEBI:CHEBI:61557; EC=3.6.1.15; Evidence={ECO:0000250|UniProtKB:Q8JUX6}; CATALYTIC ACTIVITY: Reaction=ATP + H2O = ADP + H(+) + phosphate; Xref=Rhea:RHEA:13065, ChEBI:CHEBI:15377, ChEBI:CHEBI:15378, ChEBI:CHEBI:30616, ChEBI:CHEBI:43474, ChEBI:CHEBI:456216; EC=3.6.4.13; Evidence={ECO:0000250|UniProtKB:Q8JUX6}; CATALYTIC ACTIVITY: Reaction=a ribonucleoside 5'-triphosphate + RNA(n) = diphosphate + RNA(n+1); Xref=Rhea:RHEA:21248, Rhea:RHEA-COMP:14527, Rhea:RHEA-COMP:17342, ChEBI:CHEBI:33019, ChEBI:CHEBI:61557, ChEBI:CHEBI:140395; EC=2.7.7.48; Evidence={ECO:0000255|PROSITE-ProRule:PRU00539}; CATALYTIC ACTIVITY: Reaction=4-O-(ADP-D-ribosyl)-L-aspartyl-[protein] + H2O = ADP-D-ribose + H(+) + L-aspartyl-[protein]; Xref=Rhea:RHEA:54428, Rhea:RHEA-COMP:9867, Rhea:RHEA-COMP:13832, ChEBI:CHEBI:15377, ChEBI:CHEBI:15378, ChEBI:CHEBI:29961, ChEBI:CHEBI:57967, ChEBI:CHEBI:138102; Evidence={ECO:0000269|PubMed:28150709};PhysiologicalDirection=left-to-right; Xref=Rhea:RHEA:54429; Evidence={ECO:0000269|PubMed:28150709}; CATALYTIC ACTIVITY: Reaction=5-O-(ADP-D-ribosyl)-L-glutamyl-[protein] + H2O = ADP-D-ribose + H(+) + L-glutamyl-[protein]; Xref=Rhea:RHEA:58248, Rhea:RHEA-COMP:10208, Rhea:RHEA-COMP:15089, ChEBI:CHEBI:15377, ChEBI:CHEBI:15378, ChEBI:CHEBI:29973, ChEBI:CHEBI:57967, ChEBI:CHEBI:142540; Evidence={ECO:0000269|PubMed:27440879, ECO:0000269|PubMed:28150709};PhysiologicalDirection=left-to-right; Xref=Rhea:RHEA:58249; Evidence={ECO:0000269|PubMed:27440879, ECO:0000269|PubMed:28150709}; CATALYTIC ACTIVITY: Reaction=ATP + RNA(n) = diphosphate + RNA(n)-3'-adenine ribonucleotide; Xref=Rhea:RHEA:11332, Rhea:RHEA-COMP:14527, Rhea:RHEA-COMP:17347, ChEBI:CHEBI:30616, ChEBI:CHEBI:33019, ChEBI:CHEBI:140395, ChEBI:CHEBI:173115; EC=2.7.7.19; Evidence={ECO:0000250|UniProtKB:P03317}; CATALYTIC ACTIVITY: Reaction=ADP-D-ribose 1''-phosphate + H2O = ADP-D-ribose + phosphate; Xref=Rhea:RHEA:25029, ChEBI:CHEBI:15377, ChEBI:CHEBI:43474, ChEBI:CHEBI:57967, ChEBI:CHEBI:58753; EC=3.1.3.84; Evidence={ECO:0000250|UniProtKB:P36328};PhysiologicalDirection=left-to-right; Xref=Rhea:RHEA:25030; Evidence={ECO:0000250|UniProtKB:P36328}; |
| DNA Binding | |
| EC Number | 2.1.1.-; 2.7.7.-; 3.1.3.33; 3.4.22.-; 3.6.1.15; 3.6.4.13; 3.1.3.84; 3.1.3.84; 2.7.7.19; 2.7.7.48 |
| Enzyme Function | FUNCTION: [Polyprotein P1234]: Inactive precursor of the viral replicase, which is activated by cleavages carried out by the viral protease nsP2. {ECO:0000250|UniProtKB:Q8JUX6}.; FUNCTION: [Polyprotein P123]: The early replication complex formed by the polyprotein P123 and nsP4 synthesizes the minus-strand RNAs (antigenome) (By similarity). Polyprotein P123 is a short-lived polyprotein that accumulates during early stage of infection (Probable). As soon P123 is cleaved into mature proteins, the plus-strand RNAs synthesis begins (By similarity). {ECO:0000250|UniProtKB:P03317, ECO:0000305}.; FUNCTION: [Polyprotein P123']: The early replication complex formed by the polyprotein P123' and nsP4 synthesizes minus-strand RNAs (antigenome) (Probable). Polyprotein P123' is a short-lived polyprotein that accumulates during early stage of infection (Probable). As soon P123' is cleaved into mature proteins, the plus-strand RNAs synthesis begins (Probable). {ECO:0000305}.; FUNCTION: [mRNA-capping enzyme nsP1]: Cytoplasmic capping enzyme that catalyzes two virus-specific reactions: methyltransferase and nsP1 guanylyltransferase (PubMed:26041283). mRNA-capping is necessary since all viral RNAs are synthesized in the cytoplasm, and host capping enzymes are restricted to the nucleus (Probable). The enzymatic reaction involves a covalent link between 7-methyl-GMP and nsP1, whereas eukaryotic capping enzymes form a covalent complex only with GMP (By similarity). NsP1 capping consists in the following reactions: GTP is first methylated into 7-methyl-GMP and then is covalently linked to nsP1 to form the m7GMp-nsP1 complex from which 7-methyl-GMP complex is transferred to the mRNA to create the cap structure (PubMed:26041283). NsP1 is also needed for the initiation of the minus-strand RNAs synthesis (By similarity). Probably serves as a membrane anchor for the replication complex composed of nsP1-nsP4 (By similarity). Nsp1 is needed for the initiation of the minus-strand RNAs synthesis (By similarity). Palmitoylated nsP1 is remodeling host cell cytoskeleton, and induces filopodium-like structure formation at the surface of the host cell (By similarity). {ECO:0000250|UniProtKB:P03317, ECO:0000250|UniProtKB:P08411, ECO:0000250|UniProtKB:Q8JUX6, ECO:0000269|PubMed:26041283, ECO:0000305}.; FUNCTION: [Protease nsP2]: Multifunctional protein whose N-terminus is part of the RNA polymerase complex and displays NTPase, RNA triphosphatase and helicase activities (By similarity). NTPase and RNA triphosphatase are involved in viral RNA capping and helicase keeps a check on the dsRNA replication intermediates (By similarity). The C-terminus harbors a protease that specifically cleaves the polyproteins and releases the mature proteins (By similarity). Required for the shutoff of minus-strand RNAs synthesis (By similarity). Inhibits host translation to ensure maximal viral gene expression and evade host immune response (PubMed:27318152). {ECO:0000250|UniProtKB:P03317, ECO:0000250|UniProtKB:P08411, ECO:0000250|UniProtKB:Q8JUX6, ECO:0000269|PubMed:27318152}.; FUNCTION: [Non-structural protein 3]: Seems to be essential for minus-strand RNAs and subgenomic 26S mRNAs synthesis (By similarity). Displays mono-ADP-ribosylhydrolase activity (PubMed:28150709, PubMed:27440879). ADP-ribosylation is a post-translational modification that controls various processes of the host cell and the virus probably needs to revert it for optimal viral replication (PubMed:28150709). Binds proteins of FXR family and sequesters them into the viral RNA replication complexes thereby inhibiting the formation of host stress granules on viral mRNAs (PubMed:27509095). The nsp3-FXR complexes bind viral RNAs and probably orchestrate the assembly of viral replication complexes, thanks to the ability of FXR family members to self-assemble and bind DNA (PubMed:27509095). {ECO:0000250|UniProtKB:P03317, ECO:0000269|PubMed:27440879, ECO:0000269|PubMed:27509095, ECO:0000269|PubMed:28150709}.; FUNCTION: [Non-structural protein 3']: Seems to be essential for minus-strand RNAs and subgenomic 26S mRNAs synthesis (By similarity). Displays mono-ADP-ribosylhydrolase activity (Probable). ADP-ribosylation is a post-translational modification that controls various processes of the host cell and the virus probably needs to revert it for optimal viral replication (Probable). Binds proteins of FXR family and sequesters them into the viral RNA replication complexes thereby inhibiting the formation of host stress granules on viral mRNAs (Probable). The nsp3'-FXR complexes bind viral RNAs and probably orchestrate the assembly of viral replication complexes, thanks to the ability of FXR family members to self-assemble and bind DNA (Probable). {ECO:0000250|UniProtKB:P03317, ECO:0000305|PubMed:27440879, ECO:0000305|PubMed:27509095, ECO:0000305|PubMed:28150709}.; FUNCTION: [RNA-directed RNA polymerase nsP4]: RNA dependent RNA polymerase (By similarity). Replicates genomic and antigenomic RNA by recognizing replications specific signals. The early replication complex formed by the polyproteins P123/P123' and nsP4 synthesizes minus-strand RNAs (By similarity). The late replication complex composed of fully processed nsP1-nsP4 is responsible for the production of genomic and subgenomic plus-strand RNAs (By similarity). {ECO:0000250|UniProtKB:P03317}. |
| Temperature Dependency | |
| PH Dependency | BIOPHYSICOCHEMICAL PROPERTIES: pH dependence: Optimum pH is 7 for nsP1 guanylylation. {ECO:0000269|PubMed:26041283}; |
| Pathway | |
| nucleotide Binding | NP_BIND 721..728; /note=NTP; /evidence=ECO:0000255|PROSITE-ProRule:PRU00990 |
| Features | Active site (2); Beta strand (11); Binding site (6); Chain (8); Compositional bias (2); Domain (6); Erroneous initiation (1); Helix (13); Lipidation (1); Metal binding (4); Motif (2); Mutagenesis (22); Natural variant (2); Nucleotide binding (1); Region (5); Repeat (2); Sequence conflict (3); Site (4); Turn (5) |
| Keywords | 3D-structure;ATP-binding;Eukaryotic host gene expression shutoff by virus;Eukaryotic host transcription shutoff by virus;GTP-binding;Helicase;Host cell membrane;Host cell projection;Host cytoplasm;Host cytoplasmic vesicle;Host gene expression shutoff by virus;Host membrane;Host nucleus;Host-virus interaction;Hydrolase;Inhibition of host RNA polymerase II by virus;Lipoprotein;Membrane;Metal-binding;Methyltransferase;Multifunctional enzyme;Nucleotide-binding;Nucleotidyltransferase;Palmitate;Phosphoprotein;Protease;RNA suppression of termination;RNA-binding;RNA-directed RNA polymerase;Repeat;S-adenosyl-L-methionine;Thiol protease;Transferase;Ubl conjugation;Viral RNA replication;Zinc;mRNA capping;mRNA processing |
| Interact With | |
| Induction | |
| Subcellular Location | SUBCELLULAR LOCATION: [Polyprotein P1234]: Host cytoplasmic vesicle membrane {ECO:0000305}; Peripheral membrane protein {ECO:0000305}. Note=Part of cytoplasmic vesicles, which are probably formed at the plasma membrane and internalized leading to late endosomal/lysosomal spherules containing the replication complex. {ECO:0000305}.; SUBCELLULAR LOCATION: [Polyprotein P123']: Host cytoplasmic vesicle membrane {ECO:0000305}; Peripheral membrane protein {ECO:0000305}. Note=Part of cytoplasmic vesicles, which are probably formed at the plasma membrane and internalized leading to late endosomal/lysosomal spherules containing the replication complex. {ECO:0000305}.; SUBCELLULAR LOCATION: [Polyprotein P123]: Host cytoplasmic vesicle membrane {ECO:0000305}; Peripheral membrane protein {ECO:0000305}. Note=Part of cytoplasmic vesicles, which are probably formed at the plasma membrane and internalized leading to late endosomal/lysosomal spherules containing the replication complex. {ECO:0000305}.; SUBCELLULAR LOCATION: [mRNA-capping enzyme nsP1]: Host cytoplasmic vesicle membrane {ECO:0000250|UniProtKB:P08411}; Lipid-anchor {ECO:0000250|UniProtKB:P08411}. Host cell membrane {ECO:0000250|UniProtKB:P08411}; Lipid-anchor {ECO:0000250|UniProtKB:P08411}; Cytoplasmic side {ECO:0000250|UniProtKB:P08411}. Host cell projection, host filopodium {ECO:0000250|UniProtKB:P08411}. Note=In the late phase of infection, the polyprotein is quickly cleaved before localization to cellular membranes. Then a fraction of nsP1 localizes to the inner surface of the plasma membrane and its filopodial extensions. Only the palmitoylated nsP1 localizes to the host filopodia (By similarity). NsP1 is also part of cytoplasmic vesicles, which are probably formed at the plasma membrane and internalized leading to late endosomal/lysosomal spherules containing the replication complex (By similarity). {ECO:0000250|UniProtKB:P08411}.; SUBCELLULAR LOCATION: [Protease nsP2]: Host cytoplasmic vesicle membrane {ECO:0000250|UniProtKB:P08411}; Peripheral membrane protein {ECO:0000250|UniProtKB:P08411}. Host nucleus {ECO:0000269|PubMed:17652399}. Host cytoplasm {ECO:0000269|PubMed:17652399}. Note=In the late phase of infection, the polyprotein is quickly cleaved before localization to cellular membranes. Then approximately half of nsP2 is found in the nucleus (By similarity). Shuttles between cytoplasm and nucleus (PubMed:17652399). NsP2 is also part of cytoplasmic vesicles, which are probably formed at the plasma membrane and internalized leading to late endosomal/lysosomal spherules containing the replication complex (By similarity). {ECO:0000250|UniProtKB:P08411, ECO:0000269|PubMed:17652399}.; SUBCELLULAR LOCATION: [Non-structural protein 3]: Host cytoplasmic vesicle membrane {ECO:0000250|UniProtKB:P03317}; Peripheral membrane protein {ECO:0000305}. Note=In the late phase of infection, the polyprotein is quickly cleaved before localization to cellular membranes. Then nsP3 and nsP3' form aggregates in cytoplasm (By similarity). NsP3 is also part of cytoplasmic vesicles, which are probably formed at the plasma membrane and internalized leading to late endosomal/lysosomal spherules containing the replication complex (By similarity). {ECO:0000250|UniProtKB:P03317}.; SUBCELLULAR LOCATION: [Non-structural protein 3']: Host cytoplasmic vesicle membrane {ECO:0000305}; Peripheral membrane protein {ECO:0000305}. Note=In the late phase of infection, the polyprotein is quickly cleaved before localization to cellular membranes. Then nsP3 and nsP3' form aggregates in cytoplasm (By similarity). NsP3' is also part of cytoplasmic vesicles, which are probably formed at the plasma membrane and internalized leading to late endosomal/lysosomal spherules containing the replication complex (Probable). {ECO:0000250|UniProtKB:P03317, ECO:0000305}.; SUBCELLULAR LOCATION: [RNA-directed RNA polymerase nsP4]: Host cytoplasmic vesicle membrane; Peripheral membrane protein {ECO:0000250|UniProtKB:P08411}. Note=NsP4 is part of cytoplasmic vesicles, which are probably formed at the plasma membrane and internalized leading to late endosomal/lysosomal spherules containing the replication complex. {ECO:0000250|UniProtKB:P08411}. |
| Modified Residue | |
| Post Translational Modification | PTM: [Polyprotein P1234]: Specific enzymatic cleavages in vivo yield mature proteins (By similarity). The processing of the polyprotein is temporally regulated (By similarity). In early stages (1.7 hpi), P1234 is first cleaved in trans through its nsP2 protease activity, releasing P123' and nsP4, which associate to form the early replication complex (By similarity). At the same time, P1234 is also cut at the nsP1/nsP2 site early in infection but with lower efficiency (By similarity). After replication of the viral minus-strand RNAs (4 hpi), the polyproteins are cut at the nsP1/nsP2 and nsP2/nsP3 sites very efficiently, preventing accumulation of P123' and P1234 and allowing the formation of the late replication complex (By similarity). NsP3'/nsP4 site is not cleaved anymore and P34 is produced rather than nsP4 (By similarity). {ECO:0000250|UniProtKB:P03317}.; PTM: [Polyprotein P123]: Specific enzymatic cleavages in vivo yield mature proteins (By similarity). The processing of the polyprotein is temporally regulated (By similarity). In early stages (1.7 hpi), P123 is cleaved at the nsP1/nsP2 site with low efficiency (By similarity). After replication of the viral minus-strand RNAs (4 hpi), the polyproteins are cut at the nsP1/nsP2 and nsP2/nsP3 sites very efficiently, preventing accumulation of P123 and allowing the formation of the late replication complex (By similarity). {ECO:0000250|UniProtKB:P03317}.; PTM: [Polyprotein P123']: Specific enzymatic cleavages in vivo yield mature proteins (By similarity). The processing of the polyprotein is temporally regulated (By similarity). In early stages (1.7 hpi), P123' is cleaved at the nsP1/nsP2 site with low efficiency (By similarity). After replication of the viral minus-strand RNAs (4 hpi), the polyproteins are cut at the nsP1/nsP2 and nsP2/nsP3 sites very efficiently, preventing accumulation of P123' and allowing the formation of the late replication complex (By similarity). {ECO:0000250|UniProtKB:P03317}.; PTM: [mRNA-capping enzyme nsP1]: Palmitoylated by host palmitoyltransferases ZDHHC2 and ZDHHC19. {ECO:0000250|UniProtKB:P03317}.; PTM: [Non-structural protein 3]: Phosphorylated by host on serines and threonines. {ECO:0000250|UniProtKB:P08411}.; PTM: [Non-structural protein 3']: Phosphorylated by host on serines and threonines. {ECO:0000250|UniProtKB:P08411}.; PTM: [RNA-directed RNA polymerase nsP4]: Ubiquitinated; targets the protein for rapid degradation via the ubiquitin system (By similarity). Nsp4 is present in extremely low quantities due to low frequency of translation through the amber stop-codon and the degradation by the ubiquitin pathway (By similarity). {ECO:0000250|UniProtKB:P03317}. |
| Signal Peptide | |
| Structure 3D | X-ray crystallography (4) |
| Cross Reference PDB | 2HWK; 5EZQ; 5EZS; 6BCM; |
| Mapped Pubmed ID | - |
| Motif | MOTIF 1056..1065; /note=Nuclear export signal; /evidence=ECO:0000269|PubMed:17652399; MOTIF 1179..1183; /note=Nuclear localization signal; /evidence=ECO:0000269|PubMed:17652399 |
| Gene Encoded By | |
| Mass | 277,887 |
| Kinetics | BIOPHYSICOCHEMICAL PROPERTIES: Kinetic parameters: KM=110 uM for a 12 residues substrate for nsp2 protease activity {ECO:0000269|PubMed:27030368}; |
| Metal Binding | METAL 1596; /note=Zinc; /evidence=ECO:0000250|UniProtKB:P03317; METAL 1598; /note=Zinc; /evidence=ECO:0000250|UniProtKB:P03317; METAL 1621; /note=Zinc; /evidence=ECO:0000250|UniProtKB:P03317; METAL 1639; /note=Zinc; /evidence=ECO:0000250|UniProtKB:P03317 |
| Rhea ID | RHEA:46948; RHEA:54792; RHEA:54793; RHEA:54800; RHEA:11008; RHEA:23680; RHEA:13065; RHEA:21248; RHEA:54428; RHEA:54429; RHEA:58248; RHEA:58249; RHEA:11332; RHEA:25029; RHEA:25030 |
| Cross Reference Brenda | 3.4.22.B79; |