| IED ID | IndEnz0002008196 |
| Enzyme Type ID | protease008196 |
| Protein Name |
Polyprotein nsP1234 P1234 Non-structural polyprotein Cleaved into: Polyprotein P123' P123' ; Polyprotein P123 P123 ; 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 Fragment |
| Gene Name | |
| Organism | Ross river virus (strain T48) (RRV) |
| Taxonomic Lineage | Viruses Riboviria Orthornavirae Kitrinoviricota Alsuviricetes Martellivirales Togaviridae Alphavirus (arboviruses group A) Ross River virus Ross river virus (strain T48) (RRV) |
| Enzyme Sequence | APSYRVRRTDISGHAEEAVVNAANAKGTVGDGVCRAVARKWPDSFKGAATPVGTAKLVQANGMNVIHAVGPNFSTVTEAEGDRELAAAYRAVAGIINASNIKSVAIPLLSTGVFSGGKDRVMQSLNHLFTAMDTTDADVVIYCRDKAWEKKIQEAIDRRTAVELVSEDISLESDLIRVHPDSCLVGRKGYSITDGKLHSYLEGTRFHQTAVDMAEISTLWPKLQDANEQICLYALGESMDSIRTKCPVEDADSSTPPKTVPCLCRYAMTAERVARLRMNNTKAIIVCSSFPLPKYRIEGVQKVKCDRVLIFDQTVPSLVSPRKYIPAAASTHADTVSLDSTVSTGSAWSFPSEATYETMEVVAEVHHSEPPVPPPRRRRAQVTMHHQELLEVSDMHTPIAARVEIPVYDTAVVVERVAIPCTSEYAKPIPAPRAARVVPVPAPRIQRASTYRVSPTPTPRVLRASVCSVTTSAGVEFPWAPEDLEVLTEPVHCKMREPVELPWEPEDVDIQFGDFETSDKIQFGDIDFDQFXLGRAGAYIFSSDTGPGHLQQKSVRQHALPCEMLYVHEEERTYPPALDEAREKLLQAKMQMAPTEANKSRYQSRKVENMKAVIIDRLKDGARTYLTEQSEKIPTYVSKYPRPVYSPSVEDSLQNPEVAVAACNAFLEANYPTVASYQITDEYDAYLDMVDGSESCLDRATFCPAKLRCYPKHHAYHQPQVRSAVPSPFQNTLQNVLAAATKRNCNVTQMRELPTLDSAVLNVECFKKFACNGEYWQEFKDNPIRITTENITTYVTRLKGPKAAALFAKTHNLVPLQEVPMDRFVVDMKRDVKVTPGTKHTEERPKVQVIQAAEPLATAYLCGIHRELVRRLKAVLAPNIHTLFDMSAEDFDAIIAAHFQPGDAVLETDIASFDKSQDDSLALTALMLLEDLGVDQELLDLIEAAFGEITSVHLPTGTRFKFGAMMKSGMFLTLFVNTLLNIVIACRVLREKLTNSVCAAFIGDDNIVHGVRSDPLMAERCASWVNMEVKIIDATMCEKPPYFCGGFILYDKVTGSACRVADPLKRLFKLGKPLPAGDTQDEDRRRALKDETDRWARVGLKSELEIALSSRYEVNGTGNIVRAMATLAKSLKNFKKLRGPIVHLYGGPK |
| Enzyme Length | 1149 |
| Uniprot Accession Number | P13888 |
| Absorption | |
| Active Site | |
| Activity Regulation | |
| Binding Site | BINDING 10; /note=ADP-ribose; /evidence=ECO:0000250|UniProtKB:P36328; BINDING 24; /note=ADP-ribose; /evidence=ECO:0000250|UniProtKB:Q8JUX6; BINDING 32; /note=ADP-ribose; /evidence=ECO:0000250|UniProtKB:Q8JUX6; BINDING 112; /note=ADP-ribose; /evidence=ECO:0000250|UniProtKB:P36328; BINDING 113; /note=ADP-ribose; /evidence=ECO:0000250|UniProtKB:Q8JUX6; BINDING 114; /note=ADP-ribose; /evidence=ECO:0000250|UniProtKB:P36328 |
| Calcium Binding | |
| catalytic Activity | 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:0000250|UniProtKB:Q8JUX6};PhysiologicalDirection=left-to-right; Xref=Rhea:RHEA:54429; Evidence={ECO:0000250|UniProtKB:Q8JUX6}; 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:0000250|UniProtKB:Q8JUX6};PhysiologicalDirection=left-to-right; Xref=Rhea:RHEA:58249; Evidence={ECO:0000250|UniProtKB:Q8JUX6}; 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:Q8JUX6};PhysiologicalDirection=left-to-right; Xref=Rhea:RHEA:25030; Evidence={ECO:0000250|UniProtKB:Q8JUX6}; |
| DNA Binding | |
| EC Number | 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 minus-strand RNAs (By similarity). As soon P123 is cleaved into mature proteins, the plus-strand RNAs synthesis begins (By similarity). {ECO:0000250|UniProtKB:P03317}.; FUNCTION: [Polyprotein P123']: The early replication complex formed by the polyprotein P123' and nsP4 synthesizes minus-strand RNAs (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: [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}.; 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 (By similarity). 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 (By similarity). Binds proteins of G3BP family and sequesters them into the viral RNA replication complexes thereby inhibiting the formation of host stress granules on viral mRNAs (By similarity). The nsp3-G3BP complexes bind viral RNAs and probably orchestrate the assembly of viral replication complexes, thanks to the ability of G3BP family members to self-assemble and bind DNA (By similarity). {ECO:0000250|UniProtKB:P03317, ECO:0000250|UniProtKB:Q8JUX6}.; 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 polyprotein 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 | |
| Pathway | |
| nucleotide Binding | |
| Features | Binding site (6); Chain (6); Domain (2); Metal binding (4); Modified residue (1); Motif (2); Non-terminal residue (2); Site (1) |
| Keywords | Host cytoplasmic vesicle;Host membrane;Hydrolase;Membrane;Metal-binding;Nucleotide-binding;Nucleotidyltransferase;Phosphoprotein;RNA-binding;RNA-directed RNA polymerase;Transferase;Ubl conjugation;Viral RNA replication;Zinc |
| Interact With | |
| Induction | |
| Subcellular Location | 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: [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: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: [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 | MOD_RES 344; /note=Phosphothreonine; by host; /evidence=ECO:0000250|UniProtKB:Q8JUX6 |
| 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: [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 | |
| Cross Reference PDB | - |
| Mapped Pubmed ID | - |
| Motif | MOTIF 512..515; /note=FGDF; binding to host G3BP1; /evidence=ECO:0000250|UniProtKB:P08411; MOTIF 523..526; /note=FGDF; binding to host G3BP1; /evidence=ECO:0000250|UniProtKB:P08411 |
| Gene Encoded By | |
| Mass | 127,267 |
| Kinetics | |
| Metal Binding | METAL 262; /note=Zinc; /evidence=ECO:0000250|UniProtKB:P03317; METAL 264; /note=Zinc; /evidence=ECO:0000250|UniProtKB:P03317; METAL 287; /note=Zinc; /evidence=ECO:0000250|UniProtKB:P03317; METAL 305; /note=Zinc; /evidence=ECO:0000250|UniProtKB:P03317 |
| Rhea ID | RHEA:21248; RHEA:54428; RHEA:54429; RHEA:58248; RHEA:58249; RHEA:11332; RHEA:25029; RHEA:25030 |
| Cross Reference Brenda |