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   27-MAR-2024, entry version 85.
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   Phlebovirus toscanaense.
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   Proteomes; UP000204292; Genome.
DR   GO; GO:0044165; C:host cell endoplasmic reticulum; IEA:UniProtKB-SubCell.
DR   GO; GO:0044172; C:host cell endoplasmic reticulum-Golgi intermediate compartment; IEA:UniProtKB-SubCell.
DR   GO; GO:0044177; C:host cell Golgi apparatus; IEA:UniProtKB-SubCell.
DR   GO; GO:0044423; C:virion component; IEA:UniProtKB-KW.
DR   GO; GO:0016787; F:hydrolase activity; IEA:UniProtKB-KW.
DR   GO; GO:0046872; F:metal ion binding; IEA:UniProtKB-KW.
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-dependent RNA polymerase activity; IEA:UniProtKB-KW.
DR   GO; GO:0006351; P:DNA-templated transcription; IEA:InterPro.
DR   GO; GO:0039689; P:negative stranded viral RNA replication; ISS:UniProtKB.
DR   GO; GO:0039696; P:RNA-templated viral transcription; ISS:UniProtKB.
DR   InterPro; IPR022531; L_PA-C-like.
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; L_PA-C-like; 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; Reference proteome; RNA-directed RNA polymerase;
KW   Transferase; 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
//