ID VP3_ROTSH Reviewed; 835 AA. AC A2T3S5; DT 24-MAR-2009, integrated into UniProtKB/Swiss-Prot. DT 06-MAR-2007, sequence version 1. DT 24-JAN-2024, entry version 58. DE RecName: Full=Protein VP3 {ECO:0000255|HAMAP-Rule:MF_04128}; DE Includes: DE RecName: Full=2',5'-phosphodiesterase {ECO:0000255|HAMAP-Rule:MF_04128}; DE EC=3.1.4.- {ECO:0000255|HAMAP-Rule:MF_04128, ECO:0000269|PubMed:23878220}; DE Includes: DE RecName: Full=mRNA guanylyltransferase {ECO:0000255|HAMAP-Rule:MF_04128}; DE EC=2.7.7.50 {ECO:0000255|HAMAP-Rule:MF_04128, ECO:0000269|PubMed:10603323}; DE Includes: DE RecName: Full=mRNA (guanine-N(7))-methyltransferase {ECO:0000255|HAMAP-Rule:MF_04128}; DE EC=2.1.1.56 {ECO:0000255|HAMAP-Rule:MF_04128, ECO:0000269|PubMed:10603323}; OS Rotavirus A (isolate RVA/Monkey/South Africa/SA11-H96/1958/G3P5B[2]) (RV-A) OS (Simian Agent 11 (isolate SI/South Africa/H96/58)). OC Viruses; Riboviria; Orthornavirae; Duplornaviricota; Resentoviricetes; OC Reovirales; Sedoreoviridae; Rotavirus; Rotavirus A. OX NCBI_TaxID=450149; OH NCBI_TaxID=60710; Chlorocebus pygerythrus (Vervet monkey) (Cercopithecus pygerythrus). RN [1] RP NUCLEOTIDE SEQUENCE [GENOMIC RNA]. RX PubMed=17059839; DOI=10.1016/j.virol.2006.09.024; RA Small C., Barro M., Brown T.L., Patton J.T.; RT "Genome heterogeneity of SA11 rotavirus due to reassortment with 'O' RT agent."; RL Virology 359:415-424(2007). RN [2] RP INTERACTION WITH VP2. RX PubMed=9420216; DOI=10.1128/jvi.72.1.201-208.1998; RA Zeng C.Q.-Y., Estes M.K., Charpilienne A., Cohen J.; RT "The N terminus of rotavirus VP2 is necessary for encapsidation of VP1 and RT VP3."; RL J. Virol. 72:201-208(1998). RN [3] RP FUNCTION, AND CATALYTIC ACTIVITY. RX PubMed=10603323; DOI=10.1006/viro.1999.0029; RA Chen D., Luongo C.L., Nibert M.L., Patton J.T.; RT "Rotavirus open cores catalyze 5'-capping and methylation of exogenous RNA: RT evidence that VP3 is a methyltransferase."; RL Virology 265:120-130(1999). RN [4] RP FUNCTION, DOMAIN, CATALYTIC ACTIVITY, MUTAGENESIS OF HIS-718 AND HIS-797, RP AND ACTIVE SITE. RX PubMed=23878220; DOI=10.1073/pnas.1306917110; RA Zhang R., Jha B.K., Ogden K.M., Dong B., Zhao L., Elliott R., Patton J.T., RA Silverman R.H., Weiss S.R.; RT "Homologous 2',5'-phosphodiesterases from disparate RNA viruses antagonize RT antiviral innate immunity."; RL Proc. Natl. Acad. Sci. U.S.A. 110:13114-13119(2013). RN [5] RP DOMAIN. RX PubMed=24899176; DOI=10.1128/jvi.00923-14; RA Ogden K.M., Snyder M.J., Dennis A.F., Patton J.T.; RT "Predicted structure and domain organization of rotavirus capping enzyme RT and innate immune antagonist VP3."; RL J. Virol. 88:9072-9085(2014). RN [6] RP X-RAY CRYSTALLOGRAPHY (1.39 ANGSTROMS) OF 696-835, AND DOMAIN. RX PubMed=25758703; DOI=10.1002/prot.24794; RA Brandmann T., Jinek M.; RT "Crystal structure of the C-terminal 2',5'-phosphodiesterase domain of RT group A rotavirus protein VP3."; RL Proteins 83:997-1002(2015). CC -!- FUNCTION: Multifunctional enzyme involved in mRNA capping. Catalyzes CC the formation of the 5' cap structure on the viral plus-strand CC transcripts. Specifically binds to GTP and displays guanylyltransferase CC and methyltransferase activities. Has affinity for ssRNA but not for CC dsRNA. Capping activity is non-specific and caps RNAs that initiate CC with either a G or an A residue. Together with VP1 polymerase, forms a CC VP1-VP3 complex positioned near the channels situated at each of the CC five-fold vertices of the core. Following infection, the outermost CC layer of the virus is lost, leaving a double-layered particle (DLP) CC made up of the core and VP6 shell. VP1 then catalyzes the transcription CC of fully conservative plus-strand genomic RNAs that are capped by VP3 CC and extruded through the DLP's channels into the cytoplasm where they CC function as mRNAs for translation of viral proteins. DLPs probably have CC an RNA triphosphatase activity as well, whereas open cores do not. CC {ECO:0000255|HAMAP-Rule:MF_04128, ECO:0000305|PubMed:10603323}. CC -!- FUNCTION: Counteracts the host innate immune response thanks to its CC phosphodiesterase that degrades the 5'-triphosphorylated, 2'-5' linked CC adenylate oligomers produced by the host cell IFN-inducible 2',5'- CC oligoadenylate synthetase (OAS). The host RNaseL is therefore not CC activated. {ECO:0000255|HAMAP-Rule:MF_04128, CC ECO:0000269|PubMed:23878220}. CC -!- CATALYTIC ACTIVITY: CC Reaction=a 5'-end diphospho-ribonucleoside in mRNA + GTP + H(+) = a 5'- CC end (5'-triphosphoguanosine)-(ribonucleoside) in mRNA + diphosphate; CC Xref=Rhea:RHEA:67012, Rhea:RHEA-COMP:17165, Rhea:RHEA-COMP:17166, CC ChEBI:CHEBI:15378, ChEBI:CHEBI:33019, ChEBI:CHEBI:37565, CC ChEBI:CHEBI:167616, ChEBI:CHEBI:167617; EC=2.7.7.50; CC Evidence={ECO:0000255|HAMAP-Rule:MF_04128, CC ECO:0000269|PubMed:10603323}; CC -!- CATALYTIC ACTIVITY: CC Reaction=a 5'-end (5'-triphosphoguanosine)-(ribonucleoside) in mRNA + CC S-adenosyl-L-methionine = a 5'-end (N(7)-methyl 5'- CC triphosphoguanosine)-ribonucleoside in mRNA + S-adenosyl-L- CC homocysteine; Xref=Rhea:RHEA:67008, Rhea:RHEA-COMP:17166, Rhea:RHEA- CC COMP:17167, ChEBI:CHEBI:57856, ChEBI:CHEBI:59789, ChEBI:CHEBI:156461, CC ChEBI:CHEBI:167617; EC=2.1.1.56; Evidence={ECO:0000255|HAMAP- CC Rule:MF_04128, ECO:0000269|PubMed:10603323}; CC -!- CATALYTIC ACTIVITY: CC Reaction=5'-triphosphoadenylyl-(2'->5')-adenylyl-(2'->5')-adenosine + 2 CC H2O = 2 AMP + ATP + 2 H(+); Xref=Rhea:RHEA:45964, ChEBI:CHEBI:15377, CC ChEBI:CHEBI:15378, ChEBI:CHEBI:30616, ChEBI:CHEBI:67143, CC ChEBI:CHEBI:456215; Evidence={ECO:0000255|HAMAP-Rule:MF_04128, CC ECO:0000269|PubMed:23878220}; CC -!- SUBUNIT: Interacts with VP1 (By similarity). Interacts with VP2 CC (PubMed:9420216). {ECO:0000255|HAMAP-Rule:MF_04128, CC ECO:0000269|PubMed:9420216}. CC -!- SUBCELLULAR LOCATION: Virion {ECO:0000255|HAMAP-Rule:MF_04128}. CC Note=Attached inside the inner capsid as a minor component. There are CC about 11 to 12 copies per virion. {ECO:0000255|HAMAP-Rule:MF_04128}. CC -!- DOMAIN: Contains a bipartite N7-methyltransferase domain, a 2'-O- CC methyltransferase domain and a GTase/RTPase domain. The C-terminus CC contains a phosphodiesterase domain that degrades the 5'- CC triphosphorylated, 2'-5' linked adenylate oligomers produced by the CC host cell in response to IFN stimulation. {ECO:0000255|HAMAP- CC Rule:MF_04128, ECO:0000269|PubMed:23878220, CC ECO:0000269|PubMed:24899176, ECO:0000269|PubMed:25758703}. CC -!- SIMILARITY: Belongs to the rotavirus VP3 family. {ECO:0000255|HAMAP- CC Rule:MF_04128}. 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; DQ838645; ABG75824.1; -; Genomic_RNA. DR RefSeq; YP_002302228.1; NC_011508.2. DR PDB; 5AF2; X-ray; 1.39 A; A/B/C/D=696-835. DR PDBsum; 5AF2; -. DR SMR; A2T3S5; -. DR GeneID; 7011370; -. DR KEGG; vg:7011370; -. DR Proteomes; UP000001119; Genome. DR GO; GO:0019013; C:viral nucleocapsid; IEA:UniProtKB-UniRule. DR GO; GO:0005525; F:GTP binding; IEA:UniProtKB-UniRule. DR GO; GO:0004482; F:mRNA 5'-cap (guanine-N7-)-methyltransferase activity; IEA:UniProtKB-UniRule. DR GO; GO:0004484; F:mRNA guanylyltransferase activity; IEA:UniProtKB-UniRule. DR GO; GO:0008081; F:phosphoric diester hydrolase activity; IDA:UniProtKB. DR GO; GO:0003723; F:RNA binding; IEA:UniProtKB-UniRule. DR GO; GO:0106005; P:RNA 5'-cap (guanine-N7)-methylation; IDA:UniProtKB. DR GO; GO:0039503; P:suppression by virus of host innate immune response; IEA:UniProtKB-KW. DR CDD; cd20757; capping_2-OMTase_Rotavirus; 1. DR HAMAP; MF_04124; Rota_VP3; 1. DR HAMAP; MF_04128; Rota_VP3_A; 1. DR InterPro; IPR039573; NS2A-like. DR InterPro; IPR011181; VP3_Rotav. DR Pfam; PF05213; Corona_NS2A; 1. DR Pfam; PF06929; Rotavirus_VP3; 1. DR PIRSF; PIRSF004015; LigT_rotavirus; 1. DR PROSITE; PS51589; SAM_MT56_VP3; 1. PE 1: Evidence at protein level; KW 3D-structure; GTP-binding; Host-virus interaction; Hydrolase; KW Inhibition of host innate immune response by virus; Methyltransferase; KW mRNA capping; mRNA processing; Multifunctional enzyme; Nucleotide-binding; KW Nucleotidyltransferase; Reference proteome; RNA-binding; KW S-adenosyl-L-methionine; Transferase; Viral immunoevasion; Virion. FT CHAIN 1..835 FT /note="Protein VP3" FT /id="PRO_0000367814" FT REGION 171..245 FT /note="N7-methyltransferase activity" FT /evidence="ECO:0000255|HAMAP-Rule:MF_04128, FT ECO:0000269|PubMed:24899176" FT REGION 246..428 FT /note="2'-O-methyltransferase activity" FT /evidence="ECO:0000255|HAMAP-Rule:MF_04128, FT ECO:0000269|PubMed:24899176" FT REGION 429..555 FT /note="N7-methyltransferase activity" FT /evidence="ECO:0000255|HAMAP-Rule:MF_04128, FT ECO:0000269|PubMed:24899176" FT REGION 556..693 FT /note="GTase/RTPase activity" FT /evidence="ECO:0000255|HAMAP-Rule:MF_04128, FT ECO:0000269|PubMed:24899176" FT REGION 693..835 FT /note="2'-5'-phosphodiesterase activity" FT /evidence="ECO:0000255|HAMAP-Rule:MF_04128, FT ECO:0000269|PubMed:23878220" FT ACT_SITE 718 FT /note="For 2'-5'-phosphodiesterase activity" FT /evidence="ECO:0000255|HAMAP-Rule:MF_04128, FT ECO:0000269|PubMed:23878220, ECO:0000305|PubMed:25758703" FT ACT_SITE 720 FT /note="For 2'-5'-phosphodiesterase activity" FT /evidence="ECO:0000255|HAMAP-Rule:MF_04128, FT ECO:0000305|PubMed:25758703" FT ACT_SITE 797 FT /note="For 2'-5'-phosphodiesterase activity" FT /evidence="ECO:0000255|HAMAP-Rule:MF_04128, FT ECO:0000269|PubMed:23878220, ECO:0000305|PubMed:25758703" FT ACT_SITE 799 FT /note="For 2'-5'-phosphodiesterase activity" FT /evidence="ECO:0000255|HAMAP-Rule:MF_04128, FT ECO:0000305|PubMed:25758703" FT MUTAGEN 718 FT /note="H->A: Complete loss of 2'-5'-phosphodiesterase FT activity; when associated with A-797." FT /evidence="ECO:0000269|PubMed:23878220" FT MUTAGEN 797 FT /note="H->A: Complete loss of 2'-5'-phosphodiesterase FT activity; when associated with A-718." FT /evidence="ECO:0000269|PubMed:23878220" FT STRAND 699..706 FT /evidence="ECO:0007829|PDB:5AF2" FT STRAND 714..716 FT /evidence="ECO:0007829|PDB:5AF2" FT STRAND 718..725 FT /evidence="ECO:0007829|PDB:5AF2" FT HELIX 728..730 FT /evidence="ECO:0007829|PDB:5AF2" FT HELIX 731..740 FT /evidence="ECO:0007829|PDB:5AF2" FT HELIX 742..746 FT /evidence="ECO:0007829|PDB:5AF2" FT STRAND 751..759 FT /evidence="ECO:0007829|PDB:5AF2" FT STRAND 762..766 FT /evidence="ECO:0007829|PDB:5AF2" FT HELIX 771..783 FT /evidence="ECO:0007829|PDB:5AF2" FT STRAND 787..792 FT /evidence="ECO:0007829|PDB:5AF2" FT STRAND 797..803 FT /evidence="ECO:0007829|PDB:5AF2" FT STRAND 809..814 FT /evidence="ECO:0007829|PDB:5AF2" FT STRAND 817..824 FT /evidence="ECO:0007829|PDB:5AF2" FT STRAND 829..833 FT /evidence="ECO:0007829|PDB:5AF2" SQ SEQUENCE 835 AA; 98069 MW; B761FCA189B7FBB5 CRC64; MKVLALRHSV AQVYADTQVY VHDDTKDSYE NAFLISNLTT HNILYLNYSI KTLEILNKSG IAAIALQSLE ELFTLIRCNF TYDYELDIIY LHDYSYYTNN EIRTDQHWIT KTNIEEYLLP GWKLTYVGYN GSETRGHYNF SFKCQNAATD DDLIIEYIYS EALDFQNFML KKIKERMTTS LPIARLSNRV FRDKLFPSLL KEHKNVVNVG PRNESMFTFL NYPTIKQFSN GAYLVKDTIK LKQERWLGKR ISQFDIGQYK NMLNVLTAIY YYYNLYKSKP IIYMIGSAPS YWIYDVRHYS DFFFETWDPL DTPYSSIHHK ELFFINDVKK LKDNSILYID IRTDRGNADW KKWRKTVEEQ TINNLDIAYE YLRTGKAKVC CVKMTAMDLE LPISAKLLHH PTTEIRSEFY LLLDTWDLTN IRRFIPKGVL YSFINNIITE NVFIQQPFKV KVLNDSYIVA LYALSNDFNN RSEVIKLINN QKQSLITVRI NNTFKDEPKV GFKNIYDWTF LPTDFDTKEA IITSYDGCLG LFGLSISLAS KPTGNNHLFI LSGTDKYYKL DQFANHTSIS RRSHQIRFSE SATSYSGYIF RDLSNNNFNL IGTNIENSVS GHVYNALIYY RYNYSFDLKR WIYLHSIDKV DIEGGKYYEL APIELIYACR SAKEFATLQD DLTVLRYSNE IENYINTVYS ITYADDPNYF IGIQFRNIPY KYDVKIPHLT FGVLHISDNM VPDVIDILKI MKNELFKMDI TTSYTYMLSD GIYVANVSGV LSTYFKIYNV FYKNQITFGQ SRMFIPHITL SFNNMRTVRI ETTKLQIKSI YLRKIKGDTV FDMVE //