Skip Header

Contribute Send feedback
Read comments (?) or add your own

P04298 (MCEL_VACCW) Reviewed, UniProtKB/Swiss-Prot

Last modified April 3, 2013. Version 73. Feed History...

Clusters with 100%, 90%, 50% identity | Documents (2) | Third-party data text xml rdf/xml gff fasta
to top of pageNames·Attributes·General annotation·Ontologies·Sequence annotation·Sequences·References·Cross-refs·Entry info·DocumentsCustomize order
to top of pageNames·Attributes·General annotation·Ontologies·Sequence annotation·Sequences·References·Cross-refs·Entry info·DocumentsCustomize order

Names and origin

Protein namesRecommended name:
mRNA-capping enzyme catalytic subunit
Alternative name(s):
Virus termination factor large subunit
Short name=VTF large subunit
mRNA-capping enzyme 97 kDa subunit
mRNA-capping enzyme D1 subunit
mRNA-capping enzyme large subunit

Including the following 3 domains:

  1. Polynucleotide 5'-triphosphatase
    EC=3.1.3.33
    Alternative name(s):
    mRNA 5'-triphosphatase
    Short name=TPase
  2. mRNA guanylyltransferase
    EC=2.7.7.50
    Alternative name(s):
    GTP--RNA guanylyltransferase
    Short name=GTase
  3. mRNA (guanine-N(7)-)-methyltransferase
    EC=2.1.1.56
    Alternative name(s):
    mRNA cap methyltransferase
Gene names
Ordered Locus Names:VACWR106
ORF Names:D1R
OrganismVaccinia virus (strain Western Reserve) (VACV) (Vaccinia virus (strain WR)) [Reference proteome]
Taxonomic identifier10254 [NCBI]
Taxonomic lineageViruses › dsDNA viruses, no RNA stage › Poxviridae › Chordopoxvirinae › Orthopoxvirus › Vaccinia virus
Virus hostBos taurus (Bovine) [TaxID: 9913]

Protein attributes

Sequence length844 AA.
Sequence statusComplete.
Protein existenceEvidence at protein level

General annotation (Comments)

Function

Catalytic subunit of the mRNA capping enzyme which catalyzes three enzymatic reactions: the 5' triphosphate end of the pre-mRNA is hydrolyzed to a diphosphate by RNA 5' triphosphatase; the diphosphate RNA end is capped with GMP by RNA guanylyltransferase and the GpppN cap is methylated by RNA (guanine-N7) methyltransferase. Heterodimeric mRNA capping enzyme catalyzes the linkage of a N7-methyl-guanosine moiety to the first transcribed nucleotide (cap 0 structure), whereas the polymerase associated VP39 is responsible for a second methylation at the 2'-O position of the ribose (cap 1 structure). Ref.7 Ref.8 Ref.9

The heterodimeric enzyme is also involved in early viral gene transcription termination and intermediate viral gene transcription initiation. Early gene transcription termination requires the termination factor VTF, the DNA-dependent ATPase NPH-I and the Rap94 subunit of the viral RNA polymerase, as well as the presence of a specific termination motif. Binds, together with RAP94, to the termination motif 5'-UUUUUNU-3' in the nascent early mRNA. Ref.7 Ref.8 Ref.9

Catalytic activity

A 5'-phosphopolynucleotide + H2O = a polynucleotide + phosphate.

GTP + (5')pp-Pur-mRNA = diphosphate + G(5')ppp-Pur-mRNA.

S-adenosyl-L-methionine + G(5')pppR-RNA = S-adenosyl-L-homocysteine + m7G(5')pppR-RNA.

Subunit structure

Heterodimer of a catalytic and a regulatory subunit. Intrinsic methyltransferase activity of the catalytic subunit is weak and needs to be stimulated 30- to 50-fold by the regulatory subunit, which is itself catalytically inert. Ref.3 Ref.10

Subcellular location

Virion Probable. Note: All the enzymes and other proteins required to synthesize early mRNAs are packaged within the virion core along with the DNA genome.

Domain

The N-terminus contains the triphosphatase and guanylyltransferase domains, whereas the C-terminus contains the methyltransferase domain. The N-terminus is involved in binding to the temination motif 5'-UUUUUNU-3' in the nascent mRNA. Ref.4

Sequence similarities

In the N-terminal section; belongs to the dsDNA virus mRNA guanylyltransferase family.

In the C-terminal section; belongs to the class I-like SAM-binding methyltransferase superfamily. mRNA cap 0 methyltransferase family.

Contains 1 mRNA cap 0 methyltransferase domain.

Biophysicochemical properties

Kinetic parameters:

A-570 and A-763 mutants have 3-fold and 10-fold higher Km values for GpppA.

KM=23 µM for GpppA Ref.9

Sequence annotation (Features)

Feature keyPosition(s)LengthDescriptionGraphical viewFeature identifier

Molecule processing

Chain1 – 844844mRNA-capping enzyme catalytic subunit
PRO_0000210124

Regions

Domain516 – 844329mRNA cap 0 methyltransferase
Region1 – 539539Triphosphatase-guanylyltransferase
Region549 – 5502S-adenosyl-L-methionine binding
Region569 – 5702mRNA cap binding By similarity
Region678 – 6803S-adenosyl-L-methionine binding

Sites

Active site2601N6-GMP-lysine intermediate Potential
Metal binding371Catalytic; for RNA triphosphatase activity Potential
Metal binding391Catalytic; for RNA triphosphatase activity Potential
Metal binding1921Catalytic; for RNA triphosphatase activity Potential
Metal binding1941Catalytic; for RNA triphosphatase activity Potential
Binding site5731S-adenosyl-L-methionine
Binding site5981S-adenosyl-L-methionine; via carbonyl oxygen
Binding site6201S-adenosyl-L-methionine
Binding site6821mRNA cap By similarity
Binding site7631mRNA cap By similarity
Binding site8361mRNA cap By similarity
Site771Essential for RNA triphosphatase activity
Site1071Essential for RNA triphosphatase activity
Site1261Essential for RNA triphosphatase activity
Site1591Essential for RNA triphosphatase activity
Site1611Essential for RNA triphosphatase activity
Site6071mRNA cap binding By similarity
Site6321mRNA cap binding By similarity

Natural variations

Natural variant7051G → D in strain: mutant Dts36.

Experimental info

Mutagenesis77 – 793RTK → ATA: Almost complete loss of RNA triphosphatase activity, 70% loss of guanylyltransferase activity. Ref.5
Mutagenesis1071K → A: Complete loss of triphosphatase activity, no effect on guanylyltransferase activity. Ref.6
Mutagenesis1261E → A: Complete loss of triphosphatase activity, no effect on guanylyltransferase activity. Ref.6
Mutagenesis135 – 1362KK → AA: 70% loss of RNA triphosphatase activity, 40% loss of guanylyltransferase activity.
Mutagenesis1591D → A: Complete loss of triphosphatase activity, no effect on guanylyltransferase activity. Ref.6
Mutagenesis1611K → A: Complete loss of triphosphatase activity, no effect on guanylyltransferase activity. Ref.6
Mutagenesis192 – 1943EIE → AIA: Complete loss of RNA triphosphatase activity, 50% loss of guanylyltransferase activity. Ref.5
Mutagenesis2601K → A: Complete loss of guanylyltransferase activity, no effect on RNA triphosphatase activity. Ref.5
Mutagenesis524 – 5263KIG → AIA: 50% loss of RNA triphosphatase activity and 40% loss of guanylyltransferase activity. Ref.5
Mutagenesis5701N → A: Retains substantial methyltransferase activity in vitro; lethal in vivo. Ref.9
Mutagenesis5731K → A: Complete loss of methyltransferase activity; lethal in vivo. Ref.9
Mutagenesis6071K → A: Retains substantial methyltransferase activity in vitro; lethal in vivo. Ref.9
Mutagenesis6081Y → A: Almost complete loss of methyltransferase activity; lethal in vivo. Ref.9
Mutagenesis6761D → A: Complete loss of methyltransferase activity; lethal in vivo. Ref.9
Mutagenesis6791F → A: Almost complete loss of methyltransferase activity; lethal in vivo. Ref.9
Mutagenesis6821H → A: Complete loss of methyltransferase activity; lethal in vivo. Ref.9
Mutagenesis7631E → A: Retains substantial methyltransferase activity in vitro; lethal in vivo. Ref.9

Secondary structure

1 ............................................................ 844
Helix Strand Turn

Details...

Sequences

Sequence LengthMass (Da)Tools
P04298 [UniParc].

Last modified March 20, 1987. Version 1.
Checksum: 8B9FD3DE836FA6E7

FASTA84496,734
        10         20         30         40         50         60 
MDANVVSSST IATYIDALAK NASELEQRST AYEINNELEL VFIKPPLITL TNVVNISTIQ 

        70         80         90        100        110        120 
ESFIRFTVTN KEGVKIRTKI PLSKVHGLDV KNVQLVDAID NIVWEKKSLV TENRLHKECL 

       130        140        150        160        170        180 
LRLSTEERHI FLDYKKYGSS IRLELVNLIQ AKTKNFTIDF KLKYFLGSGA QSKSSLLHAI 

       190        200        210        220        230        240 
NHPKSRPNTS LEIEFTPRDN ETVPYDELIK ELTTLSRHIF MASPENVILS PPINAPIKTF 

       250        260        270        280        290        300 
MLPKQDIVGL DLENLYAVTK TDGIPITIRV TSNGLYCYFT HLGYIIRYPV KRIIDSEVVV 

       310        320        330        340        350        360 
FGEAVKDKNW TVYLIKLIEP VNAINDRLEE SKYVESKLVD ICDRIVFKSK KYEGPFTTTS 

       370        380        390        400        410        420 
EVVDMLSTYL PKQPEGVILF YSKGPKSNID FKIKKENTID QTANVVFRYM SSEPIIFGES 

       430        440        450        460        470        480 
SIFVEYKKFS NDKGFPKEYG SGKIVLYNGV NYLNNIYCLE YINTHNEVGI KSVVVPIKFI 

       490        500        510        520        530        540 
AEFLVNGEIL KPRIDKTMKY INSEDYYGNQ HNIIVEHLRD QSIKIGDIFN EDKLSDVGHQ 

       550        560        570        580        590        600 
YANNDKFRLN PEVSYFTNKR TRGPLGILSN YVKTLLISMY CSKTFLDDSN KRKVLAIDFG 

       610        620        630        640        650        660 
NGADLEKYFY GEIALLVATD PDADAIARGN ERYNKLNSGI KTKYYKFDYI QETIRSDTFV 

       670        680        690        700        710        720 
SSVREVFYFG KFNIIDWQFA IHYSFHPRHY ATVMNNLSEL TASGGKVLIT TMDGDKLSKL 

       730        740        750        760        770        780 
TDKKTFIIHK NLPSSENYMS VEKIADDRIV VYNPSTMSTP MTEYIIKKND IVRVFNEYGF 

       790        800        810        820        830        840 
VLVDNVDFAT IIERSKKFIN GASTMEDRPS TRNFFELNRG AIKCEGLDVE DLLSYYVVYV 


FSKR

« Hide

References

« Hide 'large scale' references
[1]"Nucleotide sequence and genetic map of the 16-kb vaccinia virus HindIII D fragment."
Niles E.G., Condit R.C., Caro P., Davidson K., Matusick L., Seto J.
Virology 153:96-112(1986) [PubMed] [Europe PMC] [Abstract]
Cited for: NUCLEOTIDE SEQUENCE [GENOMIC DNA].
[2]"Sequencing of the coding region of Vaccinia-WR to an average 9-fold redundancy and an error rate of 0.16/10kb."
Esposito J.J., Frace A.M., Sammons S.A., Olsen-Rasmussen M., Osborne J., Wohlhueter R.
Submitted (FEB-2003) to the EMBL/GenBank/DDBJ databases
Cited for: NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
[3]"Interaction and mutual stabilization of the two subunits of vaccinia virus mRNA capping enzyme coexpressed in Escherichia coli."
Guo P., Moss B.
Proc. Natl. Acad. Sci. U.S.A. 87:4023-4027(1990) [PubMed] [Europe PMC] [Abstract]
Cited for: INTERACTION WITH SMALL SUBUNIT, PROTEIN SEQUENCE OF 1-20 AND 498-517.
[4]"Intrinsic RNA (guanine-7) methyltransferase activity of the vaccinia virus capping enzyme D1 subunit is stimulated by the D12 subunit. Identification of amino acid residues in the D1 protein required for subunit association and methyl group transfer."
Mao X., Shuman S.
J. Biol. Chem. 269:24472-24479(1994) [PubMed] [Europe PMC] [Abstract]
Cited for: DOMAIN METHYLTRANSFERASE.
[5]"Mutational analysis of the RNA triphosphatase component of vaccinia virus mRNA capping enzyme."
Yu L., Shuman S.
J. Virol. 70:6162-6168(1996) [PubMed] [Europe PMC] [Abstract]
Cited for: ACTIVE SITE OF TRIPHOSPHATASE ACTIVITY, MUTAGENESIS OF 77-ARG--LYS-79; 135-LYS-LYS-136; 160-PHE-LYS-161; 192-GLU--GLU-194; LYS-260 AND 524-LYS--GLY-526.
[6]"Mapping the active site of vaccinia virus RNA triphosphatase."
Gong C., Shuman S.
Virology 309:125-134(2003) [PubMed] [Europe PMC] [Abstract]
Cited for: CHARACTERIZATION OF TRIPHOSPHATASE ACTIVITY, MUTAGENESIS OF LYS-107; GLU-126; ASP-159 AND LYS-161.
[7]"Phenotypic analysis of a temperature sensitive mutant in the large subunit of the vaccinia virus mRNA capping enzyme."
Shatzer A.N., Kato S.E., Condit R.C.
Virology 375:236-252(2008) [PubMed] [Europe PMC] [Abstract]
Cited for: FUNCTION.
Strain: IHDW and mutant Dts36.
[8]"Vaccinia virus early gene transcription termination factors VTF and Rap94 interact with the U9 termination motif in the nascent RNA in a transcription ternary complex."
Christen L.A., Piacente S., Mohamed M.R., Niles E.G.
Virology 376:225-235(2008) [PubMed] [Europe PMC] [Abstract]
Cited for: FUNCTION.
[9]"Structure-function analysis of vaccinia virus mRNA cap (guanine-N7) methyltransferase."
Zheng S., Shuman S.
RNA 14:696-705(2008) [PubMed] [Europe PMC] [Abstract]
Cited for: FUNCTION, BIOPHYSICOCHEMICAL PROPERTIES, MUTAGENESIS OF ASN-570; LYS-573; LYS-607; TYR-608; ASP-676; PHE-679; HIS-682 AND GLU-763.
[10]"Structural insights into the mechanism and evolution of the vaccinia virus mRNA cap N7 methyl-transferase."
De la Pena M., Kyrieleis O.J., Cusack S.
EMBO J. 26:4913-4925(2007) [PubMed] [Europe PMC] [Abstract]
Cited for: X-RAY CRYSTALLOGRAPHY (2.7 ANGSTROMS) OF 545-844 IN COMPLEX WITH ADOHCY AND THE REGULATORY SUBUNIT.

Cross-references

Sequence databases

EMBL
GenBank
DDBJ		M15058 Genomic DNA. Translation: AAA48253.1.
AY243312 Genomic DNA. Translation: AAO89385.1.
PIRQQVZ1. A03872.
RefSeqYP_232988.1. NC_006998.1.

3D structure databases

PDBe
RCSB PDB
PDBj
EntryMethodResolution (Å)ChainPositionsPDBsum
2VDWX-ray2.70A/C/E/G545-844[»]
ProteinModelPortalP04298.
SMRP04298. Positions 545-844.
ModBaseSearch...

Protocols and materials databases

StructuralBiologyKnowledgebaseSearch...

Genome annotation databases

GeneID3707562.

Phylogenomic databases

ProtClustDBCLSP2509789.

Family and domain databases

InterProIPR019602. mRNA_cap_ATPase/GuylTrfase_vir.
IPR004971. mRNA_G-N7_MeTrfase_dom.
[Graphical view]
PfamPF10640. Pox_ATPase-GT. 1 hit.
PF03291. Pox_MCEL. 1 hit.
[Graphical view]
PROSITEPS51562. RNA_CAP0_MT. 1 hit.
[Graphical view]
ProtoNetSearch...

Other

EvolutionaryTraceP04298.

Entry information

Entry nameMCEL_VACCW
AccessionPrimary (citable) accession number: P04298
Secondary accession number(s): Q76ZS6
Entry history
Integrated into UniProtKB/Swiss-Prot: March 20, 1987
Last sequence update: March 20, 1987
Last modified: April 3, 2013
This is version 73 of the entry and version 1 of the sequence. [Complete history]
Entry statusReviewed (UniProtKB/Swiss-Prot)
Annotation programViral Protein Annotation Program

Relevant documents

PDB cross-references

Index of Protein Data Bank (PDB) cross-references

SIMILARITY comments

Index of protein domains and families

to top of pageNames·Attributes·General annotation·Ontologies·Sequence annotation·Sequences·References·Cross-refs·Entry info·Documents