L_TOSV
ID L_TOSV Reviewed; 2095 AA.
AC P37800;
DT 01-OCT-1994, integrated into UniProtKB/Swiss-Prot.
DT 01-OCT-1994, sequence version 1.
DT 03-AUG-2022, entry version 78.
DE RecName: Full=RNA-directed RNA polymerase L;
DE Short=Protein L;
DE EC=2.7.7.48 {ECO:0000250|UniProtKB:I0DF35};
DE AltName: Full=Large structural protein;
DE AltName: Full=Replicase;
DE AltName: Full=Transcriptase;
DE Includes:
DE RecName: Full=cap-snatching endonuclease;
DE EC=3.1.-.- {ECO:0000269|PubMed:31584100};
GN Name=L;
OS Toscana virus (Tos).
OC Viruses; Riboviria; Orthornavirae; Negarnaviricota; Polyploviricotina;
OC Ellioviricetes; Bunyavirales; Phenuiviridae; Phlebovirus;
OC Toscana phlebovirus.
OX NCBI_TaxID=11590;
OH NCBI_TaxID=9606; Homo sapiens (Human).
OH NCBI_TaxID=13204; Phlebotomus perniciosus.
RN [1]
RP NUCLEOTIDE SEQUENCE [GENOMIC RNA].
RC STRAIN=ISS.PHL.3;
RX PubMed=8460526; DOI=10.1016/0168-1702(93)90076-y;
RA Accardi L., Gro M.C., di Bonito P., Giorgi C.;
RT "Toscana virus genomic L segment: molecular cloning, coding strategy and
RT amino acid sequence in comparison with other negative strand RNA viruses.";
RL Virus Res. 27:119-131(1993).
RN [2]
RP REVIEW.
RX PubMed=28418734; DOI=10.1080/1040841x.2017.1307805;
RA Amroun A., Priet S., de Lamballerie X., Querat G.;
RT "Bunyaviridae RdRps: structure, motifs, and RNA synthesis machinery.";
RL Crit. Rev. Microbiol. 43:753-778(2017).
RN [3]
RP REVIEW.
RX PubMed=31948728; DOI=10.1016/j.tim.2019.12.006;
RA Olschewski S., Cusack S., Rosenthal M.;
RT "The Cap-Snatching Mechanism of Bunyaviruses.";
RL Trends Microbiol. 28:293-303(2020).
RN [4] {ECO:0007744|PDB:6QVV, ECO:0007744|PDB:6QW0, ECO:0007744|PDB:6QW5}
RP X-RAY CRYSTALLOGRAPHY (1.50 ANGSTROMS) OF 1-211, DOMAIN, CATALYTIC
RP ACTIVITY, COFACTOR, MUTAGENESIS OF HIS-78; ASP-90; ASP-113; GLU-127;
RP LYS-145 AND LYS-148, AND FUNCTION.
RC STRAIN=France AR 2005 {ECO:0000305};
RX PubMed=31584100; DOI=10.1093/nar/gkz838;
RA Jones R., Lessoued S., Meier K., Devignot S., Barata-Garcia S., Mate M.,
RA Bragagnolo G., Weber F., Rosenthal M., Reguera J.;
RT "Structure and function of the Toscana virus cap-snatching endonuclease.";
RL Nucleic Acids Res. 47:10914-10930(2019).
CC -!- FUNCTION: RNA-dependent RNA polymerase, which is responsible for the
CC replication and transcription of the viral RNA genome using antigenomic
CC RNA as an intermediate (By similarity). During transcription,
CC synthesizes subgenomic RNAs and assures their capping by a cap-
CC snatching mechanism, which involves the endonuclease activity cleaving
CC the host capped pre-mRNAs (PubMed:31584100). These short capped RNAs
CC are then used as primers for viral transcription. The 3'-end of
CC subgenomic mRNAs molecules are not polyadenylated. During replication,
CC the polymerase binds the 5' and 3' vRNA extremities at distinct sites
CC (By similarity). In turn, significant conformational changes occur in
CC the polymerase and in vRNA to initiate active RNA synthesis (By
CC similarity). As a consequence of the use of the same enzyme for both
CC transcription and replication, these mechanisms need to be well
CC coordinated (By similarity). {ECO:0000250|UniProtKB:A5HC98,
CC ECO:0000250|UniProtKB:I0DF35, ECO:0000250|UniProtKB:P27316,
CC ECO:0000269|PubMed:31584100}.
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 -!- COFACTOR:
CC Name=Mn(2+); Xref=ChEBI:CHEBI:29035;
CC Evidence={ECO:0000269|PubMed:31584100};
CC Note=For endonuclease activity. Binds 2 Mn(2+) ions in the active site
CC (PubMed:31584100). The divalent metal ions are crucial for catalytic
CC activity (PubMed:31948728, PubMed:31584100).
CC {ECO:0000269|PubMed:31584100, ECO:0000269|PubMed:31948728};
CC -!- COFACTOR:
CC Name=Mg(2+); Xref=ChEBI:CHEBI:18420;
CC Evidence={ECO:0000250|UniProtKB:P27316};
CC Name=Mn(2+); Xref=ChEBI:CHEBI:29035;
CC Evidence={ECO:0000250|UniProtKB:P27316};
CC Note=For polymerase activity. Initiation activity is stronger in the
CC presence of Mn(2+) than in the presence of Mg(2+).
CC {ECO:0000250|UniProtKB:P27316};
CC -!- SUBUNIT: Homomultimer (By similarity). Interacts with glycoprotein N;
CC this interaction allows efficient polymerase packaging into virus
CC particles (By similarity). Interacts with nucleoprotein N (By
CC similarity). {ECO:0000250|UniProtKB:P27316}.
CC -!- SUBCELLULAR LOCATION: Host Golgi apparatus
CC {ECO:0000250|UniProtKB:I0DF35}. Host endoplasmic reticulum
CC {ECO:0000250|UniProtKB:I0DF35}. Host endoplasmic reticulum-Golgi
CC intermediate compartment {ECO:0000250|UniProtKB:I0DF35}. Virion
CC {ECO:0000250|UniProtKB:P20470}.
CC -!- DOMAIN: The N-terminus contains the endonuclease activity (endoN)
CC (PubMed:31584100). The central region contains the RdRp activity (By
CC similarity). The C-terminus contains the cap-binding region (By
CC similarity). {ECO:0000250|UniProtKB:A2SZS3,
CC ECO:0000250|UniProtKB:I0DF35, ECO:0000269|PubMed:31584100}.
CC -!- MISCELLANEOUS: Classified as His(+) endonuclease since it has a
CC histidine upstream of the active site that coordinates the first
CC cation. {ECO:0000303|PubMed:31948728}.
CC -!- SIMILARITY: Belongs to the Bunyavirales RNA polymerase family.
CC {ECO:0000305}.
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DR EMBL; X68414; CAA48478.1; -; Genomic_RNA.
DR PIR; S29529; S29529.
DR PDB; 6QVV; X-ray; 2.40 A; A/B=1-211.
DR PDB; 6QW0; X-ray; 1.50 A; A/B=1-211.
DR PDB; 6QW5; X-ray; 1.99 A; A/B=1-211.
DR PDBsum; 6QVV; -.
DR PDBsum; 6QW0; -.
DR PDBsum; 6QW5; -.
DR SMR; P37800; -.
DR PRIDE; P37800; -.
DR Proteomes; UP000204292; Genome.
DR GO; GO:0001882; F:nucleoside binding; IEA:InterPro.
DR GO; GO:0000166; F:nucleotide binding; IEA:UniProtKB-KW.
DR GO; GO:0003968; F:RNA-directed 5'-3' RNA polymerase activity; IEA:UniProtKB-KW.
DR GO; GO:0039689; P:negative stranded viral RNA replication; ISS:UniProtKB.
DR GO; GO:0039696; P:RNA-templated viral transcription; ISS:UniProtKB.
DR GO; GO:0006351; P:transcription, DNA-templated; IEA:InterPro.
DR InterPro; IPR022531; DUF3770.
DR InterPro; IPR029124; L_protein_N.
DR InterPro; IPR007099; RNA-dir_pol_NSvirus.
DR InterPro; IPR014385; RNA-dir_pol_phlebovirus.
DR InterPro; IPR007322; RNA_pol_bunyavir.
DR Pfam; PF04196; Bunya_RdRp; 1.
DR Pfam; PF12603; DUF3770; 1.
DR Pfam; PF15518; L_protein_N; 1.
DR PIRSF; PIRSF000826; L_PhleboV; 1.
DR PROSITE; PS50525; RDRP_SSRNA_NEG_SEG; 1.
PE 1: Evidence at protein level;
KW 3D-structure; Host endoplasmic reticulum; Host Golgi apparatus; Hydrolase;
KW Magnesium; Manganese; Metal-binding; Nucleotide-binding;
KW Nucleotidyltransferase; RNA-directed RNA polymerase; Transferase;
KW Viral RNA replication; Virion.
FT CHAIN 1..2095
FT /note="RNA-directed RNA polymerase L"
FT /id="PRO_0000222025"
FT DOMAIN 976..1167
FT /note="RdRp catalytic"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU00539"
FT REGION 1..205
FT /note="Endonuclease"
FT /evidence="ECO:0000269|PubMed:31584100"
FT REGION 1704..1819
FT /note="Cap-binding"
FT /evidence="ECO:0000250|UniProtKB:A5HC98"
FT ACT_SITE 145
FT /note="For endonuclease activity"
FT /evidence="ECO:0000305|PubMed:31584100"
FT BINDING 78
FT /ligand="Mn(2+)"
FT /ligand_id="ChEBI:CHEBI:29035"
FT /ligand_label="1"
FT /evidence="ECO:0000269|PubMed:31584100"
FT BINDING 113
FT /ligand="Mn(2+)"
FT /ligand_id="ChEBI:CHEBI:29035"
FT /ligand_label="1"
FT /evidence="ECO:0000269|PubMed:31584100"
FT BINDING 113
FT /ligand="Mn(2+)"
FT /ligand_id="ChEBI:CHEBI:29035"
FT /ligand_label="2"
FT /evidence="ECO:0000269|PubMed:31584100"
FT BINDING 127
FT /ligand="Mn(2+)"
FT /ligand_id="ChEBI:CHEBI:29035"
FT /ligand_label="1"
FT /evidence="ECO:0000269|PubMed:31584100"
FT BINDING 1135
FT /ligand="Mg(2+)"
FT /ligand_id="ChEBI:CHEBI:18420"
FT /ligand_note="catalytic; for RdRp activity"
FT /evidence="ECO:0000250|UniProtKB:I0DF35"
FT SITE 1711
FT /note="Interaction with the cap substrate"
FT /evidence="ECO:0000250|UniProtKB:A2SZS3"
FT SITE 1715
FT /note="Interaction with the cap substrate"
FT /evidence="ECO:0000250|UniProtKB:A2SZS3"
FT SITE 1726
FT /note="Interaction with the cap substrate"
FT /evidence="ECO:0000250|UniProtKB:A2SZS3"
FT SITE 1780
FT /note="Interaction with the cap substrate"
FT /evidence="ECO:0000250|UniProtKB:A2SZS3"
FT MUTAGEN 78
FT /note="H->A: Complete loss of endonuclease activity."
FT /evidence="ECO:0000269|PubMed:31584100"
FT MUTAGEN 90
FT /note="D->A: Complete loss of endonuclease activity."
FT /evidence="ECO:0000269|PubMed:31584100"
FT MUTAGEN 113
FT /note="D->A: Complete loss of endonuclease activity."
FT /evidence="ECO:0000269|PubMed:31584100"
FT MUTAGEN 127
FT /note="E->A: Complete loss of endonuclease activity."
FT /evidence="ECO:0000269|PubMed:31584100"
FT MUTAGEN 145
FT /note="K->A: Complete loss of endonuclease activity."
FT /evidence="ECO:0000269|PubMed:31584100"
FT MUTAGEN 148
FT /note="K->A: Complete loss of endonuclease activity."
FT HELIX 4..7
FT /evidence="ECO:0007829|PDB:6QW0"
FT STRAND 15..17
FT /evidence="ECO:0007829|PDB:6QW5"
FT STRAND 21..24
FT /evidence="ECO:0007829|PDB:6QW0"
FT STRAND 35..38
FT /evidence="ECO:0007829|PDB:6QW0"
FT STRAND 45..48
FT /evidence="ECO:0007829|PDB:6QW0"
FT HELIX 50..52
FT /evidence="ECO:0007829|PDB:6QW0"
FT TURN 58..61
FT /evidence="ECO:0007829|PDB:6QW0"
FT STRAND 64..67
FT /evidence="ECO:0007829|PDB:6QW0"
FT HELIX 70..72
FT /evidence="ECO:0007829|PDB:6QW5"
FT HELIX 76..81
FT /evidence="ECO:0007829|PDB:6QW0"
FT TURN 82..85
FT /evidence="ECO:0007829|PDB:6QW0"
FT HELIX 93..97
FT /evidence="ECO:0007829|PDB:6QW0"
FT STRAND 100..103
FT /evidence="ECO:0007829|PDB:6QW5"
FT HELIX 106..109
FT /evidence="ECO:0007829|PDB:6QW0"
FT STRAND 113..117
FT /evidence="ECO:0007829|PDB:6QW0"
FT STRAND 123..130
FT /evidence="ECO:0007829|PDB:6QW0"
FT HELIX 135..156
FT /evidence="ECO:0007829|PDB:6QW0"
FT TURN 157..159
FT /evidence="ECO:0007829|PDB:6QW0"
FT STRAND 162..170
FT /evidence="ECO:0007829|PDB:6QW0"
FT STRAND 173..178
FT /evidence="ECO:0007829|PDB:6QW0"
FT HELIX 182..202
FT /evidence="ECO:0007829|PDB:6QW0"
SQ SEQUENCE 2095 AA; 238887 MW; 22FF4DAD745583B7 CRC64;
MERILKKQPA PVRALTIHPL RRYESSIYDT PIPAYVIKHS SDGVTIDIAT SELADGQSGS
TIQPFESVPA QNLTLFKHDF TFGHLADTTD KKFVEVFGVL ENRADDSDFQ SPDMIIETET
GHVYVVEFTT TMGDANSADL AARNKIAKYE IACLNRSAIK PISLYIIAVH FNGVISNLDL
SDEEVNEIVF RFRLARDIFE ELREINPALF DSDETISRLE REVNSVMSAI QIDWDTTEKK
FPSFRRELFE NFRSKEVDDE YISKIIKRCT DEALRGIERD SLYTEDITNK ERFELNSKRA
ASDIKNKMAE MMSYEFLRDT EDHKSTVQFP PWVTRTGPAG KDLEPLKSVS VEGSHPMCKI
WNKVCTNASI EKIERMHDDP VLELEYAMSG STERSVERNK YHRTVLTLSP EEREYAAVLG
VCGKRNANLG AVKEARVRSK KGFSIGHNTE RVEEFLSDSC VEDLIPTEGL YNPLSEDKSL
RLLAMGLHQP TLIHMDDETP ETLDCHLKFL SSPIGSWLQM VSIVGAELSA SVKQHVKPNQ
FIVKRLLDSA IFLLIKPTTS KGHIFVSLAV NKKFLHGELS KSSVFKQSID AGDLLVTDFV
SFKLSKITNL CKALCVLEAA SCFWAETYGF EPWKFVDQAS AVKFLDAWFM IKLSLLTMLE
DKATTEELQT MQRYVIMEGF VSLPEIPKPH KMLSKIPKVL RSELQVFLTH RLFSTMQRIS
ATPFQLHKVG GNIRWKGLFN PYSGNSIDEL QTLISCCYNG YFKNKEEDTE PSALSAMYKK
IIELEHLRPP TDTYLGYEDP IDPKMHEFSR SYLKLLCNHA KTKLRKQYGR GVMNQIENSI
VREVQSITLE RLATLKATSN FDDSWYTFKD VKDKNYTRDK LLVKMTQFAH RGKTLAIEVF
EECMSRIEEK GCMEICLFKK QQHGGLREIY VMGADERIVQ SVIEAIARAI GRFFDSDTLC
NPSNKIRIPE THGQRAKRRC GRSVWTCATS DDARKWNQGH YVTKFALMLC EFTPQEWWPL
IIRGCSMFTN KFMMMNLDFL RIIDSHKELQ IEDEFVSKLF KAYHGESVEP WISQGCTYLK
TSTGMMQGIL HFTSSLLHSL HQEFVKTTAI QLFTLKLGSD ASSKVVCDMM QGSDDSSMII
SFPSYNEKIK MRYKLVAAMC FRIKKSLGIY IGIYPSEKST PNTDFVMEYN SEFFFHSQHV
RPTIRWIAAS CSLPEVETLV ASKEEAANLL TAITEGGGSF SLAAMIQHCQ SSIHYMLMGL
GVSALFSEFS KAISKWLDPG LGFFLFDNPY SAGLSGFKYN LYRAIMNSSL KSIYSFFMKR
VKGGSQRTDG IISESCSVSP GGAIVMSSTL RWGSVEKFKR LRNRLNIPET WKEMINESPE
VLYRAPQTGT EIMLRIAEKV HSPGVVSSLS TGNAVCKVMA SSVYFLSACI FEDAGSQEYK
VVNNDKYSLM QKIIAFDQIG CNDEISQEDL LFLFPNLAEF EAFDSIIYDK GRFNVIPRAS
QREATQTRIV VFEHHSSARV APEKLVSDKW FGTRKSKIGS PGFRQEWDRL KAIVRWLRDT
PEETLDSSPF SNHIQIRNFF ARMEGRPRVI KVTGAPVKRD LGMSKIAMAI RDNFCKTGFL
QGLEDEVGHS RAMQVEKIKH YLFSVLMGPY SEEAKLEYVV KILKEEPQVI LNYNDKRSRA
NIISLLQRFI KSEIGIATLI EDMKAGVFGA FVKAQQFSQS SVNNKYYGRG IWKGVMDGYQ
VQIDIDGKEG MPSHLSGITI SNCSKTWILT QSLKAWCEDM QVYNNTDVSK ANPKANYWMY
GFKMYGSSYP YGCPIYLVRH DITNLGLLHD DDIDIKVRRN TINLFVRSKD KRPRDLHILS
YTPSDSDISS VSSKHIMEDE YFVYKGAFSV EPTRSWMLCQ PLPWSFVRPV LQVATGSRRS
PRQLDLERLR EIIRLCTESS IRNKVGTVYG QNRPEKFIEA EPIDMSEMFD MMLDEGMDDA
FEELADYLTV EEDPDYMDEV SFDDDSLNLF GPAHYKELQS LTVLAHPLMD DFVTRLVGKM
GRPQIRRLLE KNVTTRDLRE LSELLFMALD RDPSQIREEL ILGDSPTEVP DDLLG
