L_HMPVC
ID L_HMPVC Reviewed; 2005 AA.
AC Q6WB93;
DT 15-JUN-2010, integrated into UniProtKB/Swiss-Prot.
DT 05-JUL-2004, sequence version 1.
DT 03-AUG-2022, entry version 84.
DE RecName: Full=RNA-directed RNA polymerase L;
DE Short=Protein L;
DE AltName: Full=Large structural protein;
DE AltName: Full=Replicase;
DE AltName: Full=Transcriptase;
DE Includes:
DE RecName: Full=RNA-directed RNA polymerase;
DE EC=2.7.7.48 {ECO:0000250|UniProtKB:P28887};
DE Includes:
DE RecName: Full=GTP phosphohydrolase {ECO:0000250|UniProtKB:P28887};
DE EC=3.6.1.- {ECO:0000250|UniProtKB:P28887};
DE Includes:
DE RecName: Full=GDP polyribonucleotidyltransferase {ECO:0000305};
DE EC=2.7.7.88 {ECO:0000250|UniProtKB:P28887};
DE AltName: Full=PRNTase {ECO:0000305};
DE Includes:
DE RecName: Full=mRNA cap methyltransferase {ECO:0000305};
DE EC=2.1.1.375 {ECO:0000250|UniProtKB:P03523};
DE AltName: Full=mRNA (guanine-N(7)-)-methyltransferase {ECO:0000250|UniProtKB:P03523};
DE Short=G-N7-MTase {ECO:0000250|UniProtKB:P03523};
DE AltName: Full=mRNA (nucleoside-2'-O-)-methyltransferase {ECO:0000250|UniProtKB:P03523};
DE Short=N1-2'-O-MTase {ECO:0000250|UniProtKB:P03523};
GN Name=L;
OS Human metapneumovirus (strain CAN97-83) (HMPV).
OC Viruses; Riboviria; Orthornavirae; Negarnaviricota; Haploviricotina;
OC Monjiviricetes; Mononegavirales; Pneumoviridae; Metapneumovirus.
OX NCBI_TaxID=694067;
OH NCBI_TaxID=9606; Homo sapiens (Human).
RN [1]
RP NUCLEOTIDE SEQUENCE [GENOMIC RNA].
RX PubMed=14592754; DOI=10.1016/s0042-6822(03)00528-2;
RA Biacchesi S., Skiadopoulos M.H., Boivin G., Hanson C.T., Murphy B.R.,
RA Collins P.L., Buchholz U.J.;
RT "Genetic diversity between human metapneumovirus subgroups.";
RL Virology 315:1-9(2003).
RN [2]
RP NUCLEOTIDE SEQUENCE [GENOMIC RNA].
RX PubMed=16306583; DOI=10.1128/jvi.79.24.15114-15122.2005;
RA Pham Q.N., Biacchesi S., Skiadopoulos M.H., Murphy B.R., Collins P.L.,
RA Buchholz U.J.;
RT "Chimeric recombinant human metapneumoviruses with the nucleoprotein or
RT phosphoprotein open reading frame replaced by that of avian metapneumovirus
RT exhibit improved growth in vitro and attenuation in vivo.";
RL J. Virol. 79:15114-15122(2005).
RN [3]
RP CATALYTIC ACTIVITY, ACTIVE SITE, AND MUTAGENESIS OF GLY-1696; GLY-1700 AND
RP ASP-1755.
RC STRAIN=NL;
RX PubMed=25056882; DOI=10.1128/jvi.00876-14;
RA Zhang Y., Wei Y., Zhang X., Cai H., Niewiesk S., Li J.;
RT "Rational design of human metapneumovirus live attenuated vaccine
RT candidates by inhibiting viral mRNA cap methyltransferase.";
RL J. Virol. 88:11411-11429(2014).
RN [4] {ECO:0007744|PDB:6U5O}
RP STRUCTURE BY ELECTRON MICROSCOPY (3.70 ANGSTROMS), DOMAIN, AND MUTAGENESIS
RP OF ASP-745.
RX PubMed=31698413; DOI=10.1038/s41586-019-1759-1;
RA Pan J., Qian X., Lattmann S., El Sahili A., Yeo T.H., Jia H., Cressey T.,
RA Ludeke B., Noton S., Kalocsay M., Fearns R., Lescar J.;
RT "Structure of the human metapneumovirus polymerase phosphoprotein
RT complex.";
RL Nature 577:275-279(2020).
CC -!- FUNCTION: Responsible for RNA synthesis (replicase and transcriptase),
CC cap addition, and cap methylation. Performs also the polyadenylation of
CC subgenomic mRNAs by a stuttering mechanism at a slipery stop site
CC present at the end of viral genes. The template is composed of the
CC viral RNA tightly encapsidated by the nucleoprotein (N). The viral
CC polymerase binds to the genomic RNA at two differents sites in the 3'
CC leader promoter thereby initiating either genome replication or mRNA
CC transcription. In the transcription mode, the polymerase performs the
CC sequential transcription of all mRNAs using a termination-reinitiation
CC mechanism responding to gene start and gene end signals. Some
CC polymerase disengage from the template at each gene junction, resulting
CC in a decreasing abundance of transcripts from the 3' to the 5' end of
CC the genome. The first gene is the most transcribed, and the last the
CC least transcribed. Needs as cofactors the phosphoprotein for
CC processivity and the M2-1 anti-termination protein. Polyribonucleotidyl
CC transferase (PRNTase) adds the cap structure when the nascent RNA chain
CC length has reached few nucleotides (By similarity). Ribose 2'-O
CC methylation of viral mRNA cap precedes and facilitates subsequent
CC guanine-N-7 methylation (By similarity). In the replication mode, the
CC polymerase replicates the whole viral genome without recognizing the
CC gene end transcriptional signals. The ability of the polymerase to
CC override the gene end signals as it is producing the antigenome is
CC probably due to replicative RNA becoming encapsidated with
CC nucleoprotein as it is synthesized (By similarity).
CC {ECO:0000250|UniProtKB:P03523, ECO:0000250|UniProtKB:P28887}.
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 -!- CATALYTIC ACTIVITY:
CC Reaction=GTP + H2O = GDP + H(+) + phosphate; Xref=Rhea:RHEA:19669,
CC ChEBI:CHEBI:15377, ChEBI:CHEBI:15378, ChEBI:CHEBI:37565,
CC ChEBI:CHEBI:43474, ChEBI:CHEBI:58189;
CC Evidence={ECO:0000250|UniProtKB:P28887};
CC -!- CATALYTIC ACTIVITY:
CC Reaction=a 5'-end triphospho-adenylyl-adenylyl-cytidylyl-adenosine in
CC mRNA + GDP + H(+) = a 5'-end (5'-triphosphoguanosine)-adenylyl-
CC adenylyl-cytidylyl-adenosine in mRNA + diphosphate;
CC Xref=Rhea:RHEA:65436, Rhea:RHEA-COMP:16797, Rhea:RHEA-COMP:16799,
CC ChEBI:CHEBI:15378, ChEBI:CHEBI:33019, ChEBI:CHEBI:58189,
CC ChEBI:CHEBI:156484, ChEBI:CHEBI:156503; EC=2.7.7.88;
CC Evidence={ECO:0000250|UniProtKB:P28887};
CC -!- CATALYTIC ACTIVITY:
CC Reaction=a 5'-end (5'-triphosphoguanosine)-adenylyl-adenylyl-cytidylyl-
CC adenosine in mRNA + 2 S-adenosyl-L-methionine = a 5'-end (N(7)-methyl
CC 5'-triphosphoguanosine)-(2'-O-methyladenylyl)-adenylyl-cytidylyl-
CC adenosine in mRNA + H(+) + 2 S-adenosyl-L-homocysteine;
CC Xref=Rhea:RHEA:65376, Rhea:RHEA-COMP:16797, Rhea:RHEA-COMP:16798,
CC ChEBI:CHEBI:15378, ChEBI:CHEBI:57856, ChEBI:CHEBI:59789,
CC ChEBI:CHEBI:156483, ChEBI:CHEBI:156484; EC=2.1.1.375;
CC Evidence={ECO:0000269|PubMed:25056882};
CC -!- CATALYTIC ACTIVITY:
CC Reaction=a 5'-end (5'-triphosphoguanosine)-adenylyl-adenylyl-cytidylyl-
CC adenosine in mRNA + S-adenosyl-L-methionine = a 5'-end (5'-
CC triphosphoguanosine)-(2'-O-methyladenylyl)-adenylyl-cytidylyl-
CC adenosine in mRNA + H(+) + S-adenosyl-L-homocysteine;
CC Xref=Rhea:RHEA:65380, Rhea:RHEA-COMP:16797, Rhea:RHEA-COMP:16801,
CC ChEBI:CHEBI:15378, ChEBI:CHEBI:57856, ChEBI:CHEBI:59789,
CC ChEBI:CHEBI:156482, ChEBI:CHEBI:156484;
CC Evidence={ECO:0000250|UniProtKB:P03523};
CC -!- CATALYTIC ACTIVITY:
CC Reaction=a 5'-end (5'-triphosphoguanosine)-(2'-O-methyladenylyl)-
CC adenylyl-cytidylyl-adenosine in mRNA + S-adenosyl-L-methionine = a
CC 5'-end (N(7)-methyl 5'-triphosphoguanosine)-(2'-O-methyladenylyl)-
CC adenylyl-cytidylyl-adenosine in mRNA + S-adenosyl-L-homocysteine;
CC Xref=Rhea:RHEA:65440, Rhea:RHEA-COMP:16798, Rhea:RHEA-COMP:16801,
CC ChEBI:CHEBI:57856, ChEBI:CHEBI:59789, ChEBI:CHEBI:156482,
CC ChEBI:CHEBI:156483; Evidence={ECO:0000250|UniProtKB:P03523};
CC -!- COFACTOR:
CC Name=Mg(2+); Xref=ChEBI:CHEBI:18420;
CC Evidence={ECO:0000250|UniProtKB:P28887};
CC Note=For RNA-directed RNA polymerase activity. Mn(2+) can stimulate de
CC novo initiation but it is inefficient at supporting elongation of de
CC novo initiated RNA. {ECO:0000250|UniProtKB:P28887};
CC -!- SUBUNIT: Interacts with the phosphoprotein (via C-terminus); the
CC association of P and L forms the polymerase complex.
CC {ECO:0000250|UniProtKB:P28887}.
CC -!- SUBCELLULAR LOCATION: Virion {ECO:0000250|UniProtKB:P28887}. Host
CC cytoplasm {ECO:0000250|UniProtKB:P28887}. Note=Localizes in cytoplasmic
CC inclusion bodies. {ECO:0000250|UniProtKB:P28887}.
CC -!- DOMAIN: Contains an RNA-dependent RNA polymerase (RdRp) domain, a
CC polyribonucleotidyl transferase (PRNTase or capping) domain and a
CC methyltransferase (MTase) domain. {ECO:0000250|UniProtKB:P28887}.
CC -!- SIMILARITY: Belongs to the paramyxovirus L protein family.
CC {ECO:0000305}.
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; AY297749; AAQ67700.1; -; Genomic_RNA.
DR RefSeq; YP_012613.1; NC_004148.2.
DR PDB; 4UCJ; X-ray; 3.26 A; A/B=1600-2005.
DR PDB; 6U5O; EM; 3.70 A; L=1-2005.
DR PDBsum; 4UCJ; -.
DR PDBsum; 6U5O; -.
DR SMR; Q6WB93; -.
DR IntAct; Q6WB93; 1.
DR PRIDE; Q6WB93; -.
DR Proteomes; UP000001398; Genome.
DR GO; GO:0030430; C:host cell cytoplasm; IEA:UniProtKB-SubCell.
DR GO; GO:0005524; F:ATP binding; IEA:UniProtKB-KW.
DR GO; GO:0003924; F:GTPase activity; IEA:RHEA.
DR GO; GO:0046872; F:metal ion binding; IEA:UniProtKB-KW.
DR GO; GO:0004482; F:mRNA (guanine-N7-)-methyltransferase activity; IEA:InterPro.
DR GO; GO:0003968; F:RNA-directed 5'-3' RNA polymerase activity; IEA:UniProtKB-KW.
DR InterPro; IPR039736; L_poly_C.
DR InterPro; IPR026890; Mononeg_mRNAcap.
DR InterPro; IPR014023; Mononeg_RNA_pol_cat.
DR InterPro; IPR025786; Mononega_L_MeTrfase.
DR Pfam; PF14318; Mononeg_mRNAcap; 1.
DR Pfam; PF00946; Mononeg_RNA_pol; 1.
DR TIGRFAMs; TIGR04198; paramyx_RNAcap; 1.
DR PROSITE; PS50526; RDRP_SSRNA_NEG_NONSEG; 1.
DR PROSITE; PS51590; SAM_MT_MNV_L; 1.
PE 1: Evidence at protein level;
KW 3D-structure; ATP-binding; Host cytoplasm; Hydrolase; Magnesium;
KW Metal-binding; Methyltransferase; mRNA capping; mRNA processing;
KW Multifunctional enzyme; Nucleotide-binding; Nucleotidyltransferase;
KW Reference proteome; RNA-directed RNA polymerase; S-adenosyl-L-methionine;
KW Transferase; Viral RNA replication; Virion.
FT CHAIN 1..2005
FT /note="RNA-directed RNA polymerase L"
FT /id="PRO_0000394810"
FT DOMAIN 628..811
FT /note="RdRp catalytic"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU00539"
FT DOMAIN 1662..1852
FT /note="Mononegavirus-type SAM-dependent 2'-O-MTase"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU00923"
FT REGION 902..1379
FT /note="GDP polyribonucleotidyltransferase"
FT /evidence="ECO:0000250|UniProtKB:P28887"
FT ACT_SITE 1263
FT /note="Nucleophile; for GDP polyribonucleotidyltransferase
FT activity"
FT /evidence="ECO:0000250|UniProtKB:P03523"
FT ACT_SITE 1673
FT /note="For mRNA (nucleoside-2'-O-)-methyltransferase
FT activity"
FT /evidence="ECO:0000269|PubMed:25056882"
FT ACT_SITE 1779
FT /note="For mRNA (nucleoside-2'-O-)-methyltransferase
FT activity"
FT /evidence="ECO:0000269|PubMed:25056882"
FT ACT_SITE 1817
FT /note="For mRNA (nucleoside-2'-O-)-methyltransferase
FT activity"
FT /evidence="ECO:0000269|PubMed:25056882"
FT ACT_SITE 1848
FT /note="For mRNA (nucleoside-2'-O-)-methyltransferase
FT activity"
FT /evidence="ECO:0000269|PubMed:25056882"
FT BINDING 635
FT /ligand="Mg(2+)"
FT /ligand_id="ChEBI:CHEBI:18420"
FT /ligand_note="catalytic; for RNA-directed RNA polymerase
FT activity"
FT /evidence="ECO:0000250|UniProtKB:P28887"
FT BINDING 745
FT /ligand="Mg(2+)"
FT /ligand_id="ChEBI:CHEBI:18420"
FT /ligand_note="catalytic; for RNA-directed RNA polymerase
FT activity"
FT /evidence="ECO:0000305|PubMed:31698413"
FT BINDING 1696..1700
FT /ligand="substrate"
FT /ligand_note="for mRNA (nucleoside-2'-O-)-methyltransferase
FT activity"
FT /evidence="ECO:0000269|PubMed:25056882"
FT MUTAGEN 745
FT /note="D->A: Complete loss of polymerase activity."
FT /evidence="ECO:0000269|PubMed:31698413"
FT MUTAGEN 1696
FT /note="G->A: Impaired cell-cell spread and/or viral
FT replication. Loss of 2'-O methylation activity but not G-N-
FT 7 methylation activity."
FT /evidence="ECO:0000269|PubMed:25056882"
FT MUTAGEN 1700
FT /note="G->A: Impaired cell-cell spread and/or viral
FT replication. 70-80% reduction in N-protein expression. Loss
FT of 2'-O methylation activity but not G-N-7 methylation
FT activity."
FT /evidence="ECO:0000269|PubMed:25056882"
FT MUTAGEN 1755
FT /note="D->A: Impaired cell-cell spread and/or viral
FT replication. 50-70% reduction in N-protein expression. Loss
FT of 2'-O methylation activity but not G-N-7 methylation
FT activity."
FT /evidence="ECO:0000269|PubMed:25056882"
SQ SEQUENCE 2005 AA; 230760 MW; C87A1F997DBBBD2D CRC64;
MDPLNESTVN VYLPDSYLKG VISFSETNAI GSCLLKRPYL KNDNTAKVAI ENPVIEHVRL
KNAVNSKMKI SDYKVVEPVN MQHEIMKNVH SCELTLLKQF LTRSKNISTL KLNMICDWLQ
LKSTSDDTSI LSFIDVEFIP SWVSNWFSNW YNLNKLILEF RREEVIRTGS ILCRSLGKLV
FIVSSYGCIV KSNKSKRVSF FTYNQLLTWK DVMLSRFNAN FCIWVSNSLN ENQEGLGLRS
NLQGMLTNKL YETVDYMLSL CCNEGFSLVK EFEGFIMSEI LRITEHAQFS TRFRNTLLNG
LTDQLTKLKN KNRLRVHSTV LENNDYPMYE VVLKLLGDTL RCIKLLINKN LENAAELYYI
FRIFGHPMVD ERDAMDAVKL NNEITKILRL ESLTELRGAF ILRIIKGFVD NNKRWPKIKN
LKVLSKRWTM YFKAKNYPSQ LELSEQDFLE LAAIQFEQEF SVPEKTNLEM VLNDKAISPP
KRLIWSVYPK NYLPETIKNR YLEETFNASD SLKTRRVLEY YLKDNKFDQK ELKSYVVRQE
YLNDKEHIVS LTGKERELSV GRMFAMQPGK QRQIQILAEK LLADNIVPFF PETLTKYGDL
DLQRIMEIKS ELSSIKTRRN DSYNNYIARA SIVTDLSKFN QAFRYETTAI CADVADELHG
TQSLFCWLHL IVPMTTMICA YRHAPPETKG EYDIDKIEEQ SGLYRYHMGG IEGWCQKLWT
MEAISLLDVV SVKTRCQMTS LLNGDNQSID VSKPVKLSEG LDEVKADYRL AVKMLKEIRD
AYRNIGHKLK EGETYISRDL QFISKVIQSE GVMHPTPIKK VLRVGPWINT ILDDIKTSAE
SIGSLCQELE FRGESIIVSL ILRNFWLYNL YMHESKQHPL AGKQLFKQLN KTLTSVQRFF
EIKRENEVVD LWMNIPMQFG GGDPVVFYRS FYRRTPDFLT EAISHVDILL KISANIKNET
KVSFFKALLS IEKNERATLT TLMRDPQAVG SERQAKVTSD INRTAVTSIL SLSPNQLFSD
SAIHYSRNEE EVGIIAENIT PVYPHGLRVL YESLPFHKAE KVVNMISGTK SITNLLQRTS
AINGEDIDRA VSMMLENLGL LSRILSVVVD SIEIPIKSNG RLICCQISRT LRETSWNNME
IVGVTSPSIT TCMDVIYATS SHLKGIIIEK FSTDRTTRGQ RGPKSPWVGS STQEKKLVPV
YNRQILSKQQ REQLEAIGKM RWVYKGTPGL RRLLNKICLG SLGISYKCVK PLLPRFMSVN
FLHRLSVSSR PMEFPASVPA YRTTNYHFDT SPINQALSER FGNEDINLVF QNAISCGISI
MSVVEQLTGR SPKQLVLIPQ LEEIDIMPPP VFQGKFNYKL VDKITSDQHI FSPDKIDMLT
LGKMLMPTIK GQKTDQFLNK RENYFHGNNL IESLSAALAC HWCGILTEQC IENNIFKKDW
GDGFISDHAF MDFKIFLCVF KTKLLCSWGS QGKNIKDEDI VDESIDKLLR IDNTFWRMFS
KVMFEPKVKK RIMLYDVKFL SLVGYIGFKN WFIEQLRSAE LHEIPWIVNA EGDLVEIKSI
KIYLQLIEQS LFLRITVLNY TDMAHALTRL IRKKLMCDNA LLTPISSPMV NLTQVIDPTT
QLDYFPKITF ERLKNYDTSS NYAKGKLTRN YMILLPWQHV NRYNFVFSST GCKVSLKTCI
GKLMKDLNPK VLYFIGEGAG NWMARTACEY PDIKFVYRSL KDDLDHHYPL EYQRVIGELS
RIIDSGEGLS METTDATQKT HWDLIHRVSK DALLITLCDA EFKDRDDFFK MVILWRKHVL
SCRICTTYGT DLYLFAKYHA KDCNVKLPFF VRSVATFIMQ GSKLSGSECY ILLTLGHHNS
LPCHGEIQNS KMKIAVCNDF YAAKKLDNKS IEANCKSLLS GLRIPINKKE LDRQRRLLTL
QSNHSSVATV GGSKIIESKW LTNKASTIID WLEHILNSPK GELNYDFFEA LENTYPNMIK
LIDNLGNAEI KKLIKVTGYM LVSKK
