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P04298

- MCEL_VACCW

UniProt

P04298 - MCEL_VACCW

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Protein

mRNA-capping enzyme catalytic subunit

Gene

VACWR106

Organism
Vaccinia virus (strain Western Reserve) (VACV) (Vaccinia virus (strain WR))
Status
Reviewed - Annotation score: 5 out of 5- Experimental evidence at protein leveli

Functioni

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).
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.

Catalytic activityi

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.PROSITE-ProRule annotation

Kineticsi

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

  1. KM=23 µM for GpppA1 Publication

Sites

Feature keyPosition(s)LengthDescriptionGraphical viewFeature identifierActions
Metal bindingi37 – 371Catalytic; for RNA triphosphatase activitySequence Analysis
Metal bindingi39 – 391Catalytic; for RNA triphosphatase activitySequence Analysis
Sitei77 – 771Essential for RNA triphosphatase activity
Sitei107 – 1071Essential for RNA triphosphatase activity
Sitei126 – 1261Essential for RNA triphosphatase activity
Sitei159 – 1591Essential for RNA triphosphatase activity
Sitei161 – 1611Essential for RNA triphosphatase activity
Metal bindingi192 – 1921Catalytic; for RNA triphosphatase activitySequence Analysis
Metal bindingi194 – 1941Catalytic; for RNA triphosphatase activitySequence Analysis
Active sitei260 – 2601N6-GMP-lysine intermediateSequence Analysis
Binding sitei573 – 5731S-adenosyl-L-methionine
Binding sitei598 – 5981S-adenosyl-L-methionine; via carbonyl oxygen
Sitei607 – 6071mRNA cap bindingPROSITE-ProRule annotation
Binding sitei620 – 6201S-adenosyl-L-methionine
Sitei632 – 6321mRNA cap bindingPROSITE-ProRule annotation
Binding sitei682 – 6821mRNA capPROSITE-ProRule annotation
Binding sitei763 – 7631mRNA capPROSITE-ProRule annotation
Binding sitei836 – 8361mRNA capPROSITE-ProRule annotation

GO - Molecular functioni

  1. GTP binding Source: UniProtKB-KW
  2. metal ion binding Source: UniProtKB-KW
  3. mRNA (guanine-N7-)-methyltransferase activity Source: UniProtKB-EC
  4. mRNA guanylyltransferase activity Source: UniProtKB-EC
  5. polynucleotide 5'-phosphatase activity Source: UniProtKB-EC
  6. RNA binding Source: UniProtKB-KW

GO - Biological processi

  1. DNA-templated transcription, termination Source: UniProtKB-KW
  2. regulation of transcription, DNA-templated Source: UniProtKB-KW
Complete GO annotation...

Keywords - Molecular functioni

Hydrolase, Methyltransferase, Nucleotidyltransferase, Transferase

Keywords - Biological processi

mRNA capping, mRNA processing, Transcription, Transcription regulation, Transcription termination

Keywords - Ligandi

GTP-binding, Metal-binding, Nucleotide-binding, RNA-binding, S-adenosyl-L-methionine

Names & Taxonomyi

Protein namesi
Recommended 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:
Polynucleotide 5'-triphosphatase (EC:3.1.3.33)
Alternative name(s):
mRNA 5'-triphosphatase
Short name:
TPase
mRNA guanylyltransferase (EC:2.7.7.50)
Alternative name(s):
GTP--RNA guanylyltransferase
Short name:
GTase
mRNA (guanine-N(7)-)-methyltransferase (EC:2.1.1.56)
Alternative name(s):
mRNA cap methyltransferase
Gene namesi
Ordered Locus Names:VACWR106
ORF Names:D1R
OrganismiVaccinia virus (strain Western Reserve) (VACV) (Vaccinia virus (strain WR))
Taxonomic identifieri10254 [NCBI]
Taxonomic lineageiVirusesdsDNA viruses, no RNA stagePoxviridaeChordopoxvirinaeOrthopoxvirusVaccinia virus
Virus hostiBos taurus (Bovine) [TaxID: 9913]
ProteomesiUP000000344: Genome

Subcellular locationi

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

GO - Cellular componenti

  1. virion Source: UniProtKB-KW
Complete GO annotation...

Keywords - Cellular componenti

Virion

Pathology & Biotechi

Mutagenesis

Feature keyPosition(s)LengthDescriptionGraphical viewFeature identifierActions
Mutagenesisi77 – 793RTK → ATA: Almost complete loss of RNA triphosphatase activity, 70% loss of guanylyltransferase activity. 1 Publication
Mutagenesisi107 – 1071K → A: Complete loss of triphosphatase activity, no effect on guanylyltransferase activity. 1 Publication
Mutagenesisi126 – 1261E → A: Complete loss of triphosphatase activity, no effect on guanylyltransferase activity. 1 Publication
Mutagenesisi135 – 1362KK → AA: 70% loss of RNA triphosphatase activity, 40% loss of guanylyltransferase activity. 1 Publication
Mutagenesisi159 – 1591D → A: Complete loss of triphosphatase activity, no effect on guanylyltransferase activity. 1 Publication
Mutagenesisi161 – 1611K → A: Complete loss of triphosphatase activity, no effect on guanylyltransferase activity. 1 Publication
Mutagenesisi192 – 1943EIE → AIA: Complete loss of RNA triphosphatase activity, 50% loss of guanylyltransferase activity. 1 Publication
Mutagenesisi260 – 2601K → A: Complete loss of guanylyltransferase activity, no effect on RNA triphosphatase activity. 1 Publication
Mutagenesisi524 – 5263KIG → AIA: 50% loss of RNA triphosphatase activity and 40% loss of guanylyltransferase activity. 1 Publication
Mutagenesisi570 – 5701N → A: Retains substantial methyltransferase activity in vitro; lethal in vivo. 1 Publication
Mutagenesisi573 – 5731K → A: Complete loss of methyltransferase activity; lethal in vivo. 1 Publication
Mutagenesisi607 – 6071K → A: Retains substantial methyltransferase activity in vitro; lethal in vivo. 1 Publication
Mutagenesisi608 – 6081Y → A: Almost complete loss of methyltransferase activity; lethal in vivo. 1 Publication
Mutagenesisi676 – 6761D → A: Complete loss of methyltransferase activity; lethal in vivo. 1 Publication
Mutagenesisi679 – 6791F → A: Almost complete loss of methyltransferase activity; lethal in vivo. 1 Publication
Mutagenesisi682 – 6821H → A: Complete loss of methyltransferase activity; lethal in vivo. 1 Publication
Mutagenesisi763 – 7631E → A: Retains substantial methyltransferase activity in vitro; lethal in vivo. 1 Publication

PTM / Processingi

Molecule processing

Feature keyPosition(s)LengthDescriptionGraphical viewFeature identifierActions
Chaini1 – 844844mRNA-capping enzyme catalytic subunitPRO_0000210124Add
BLAST

Interactioni

Subunit structurei

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.1 Publication

Protein-protein interaction databases

DIPiDIP-60664N.

Structurei

Secondary structure

1
844
Legend: HelixTurnBeta strand
Show more details
Feature keyPosition(s)LengthDescriptionGraphical viewFeature identifierActions
Helixi3 – 53
Helixi11 – 2414
Beta strandi36 – 449
Helixi48 – 547
Beta strandi58 – 6912
Beta strandi71 – 788
Helixi82 – 843
Beta strandi87 – 893
Helixi92 – 976
Beta strandi103 – 11614
Beta strandi119 – 13012
Helixi134 – 1407
Beta strandi142 – 15211
Beta strandi154 – 17017
Helixi175 – 1817
Beta strandi182 – 1854
Beta strandi188 – 1958
Beta strandi198 – 2003
Helixi205 – 22016
Beta strandi226 – 2294
Beta strandi236 – 2416
Helixi244 – 2474
Beta strandi255 – 2595
Beta strandi262 – 27110
Beta strandi274 – 2796
Turni280 – 2823
Beta strandi283 – 2886
Beta strandi298 – 31821
Helixi327 – 33711
Turni338 – 3403
Beta strandi343 – 3497
Helixi359 – 36911
Turni370 – 3723
Beta strandi377 – 3848
Helixi385 – 3884
Beta strandi389 – 3935
Beta strandi398 – 4058
Beta strandi422 – 43211
Beta strandi435 – 44511
Beta strandi447 – 4537
Turni454 – 4574
Beta strandi458 – 4658
Helixi466 – 4683
Beta strandi470 – 48415
Beta strandi489 – 4924
Helixi494 – 5029
Beta strandi508 – 5103
Helixi511 – 52212
Helixi525 – 5273
Helixi563 – 58018
Turni583 – 5853
Beta strandi593 – 5964
Turni600 – 6045
Helixi605 – 6106
Beta strandi614 – 6218
Helixi623 – 63614
Beta strandi646 – 6516
Beta strandi656 – 6583
Helixi659 – 6646
Beta strandi672 – 6798
Helixi681 – 6833
Turni687 – 6893
Helixi690 – 70011
Beta strandi701 – 71212
Helixi714 – 7174
Beta strandi724 – 7274
Beta strandi730 – 7323
Turni734 – 7363
Beta strandi737 – 7415
Beta strandi743 – 7453
Beta strandi748 – 7525
Turni754 – 7563
Beta strandi757 – 7593
Beta strandi761 – 7644
Helixi768 – 77710
Beta strandi780 – 7878
Helixi788 – 7936
Helixi796 – 8005
Helixi802 – 8054
Helixi809 – 82315
Beta strandi826 – 8283
Helixi829 – 8335
Beta strandi836 – 8438

3D structure databases

Select the link destinations:
PDBe
RCSB PDB
PDBj
Links Updated
EntryMethodResolution (Å)ChainPositionsPDBsum
2VDWX-ray2.70A/C/E/G545-844[»]
4CKBX-ray2.80A/D1-844[»]
4CKCX-ray2.90A/D1-844[»]
4CKEX-ray2.90A/D1-844[»]
ProteinModelPortaliP04298.
SMRiP04298. Positions 545-844.
ModBaseiSearch...
MobiDBiSearch...

Miscellaneous databases

EvolutionaryTraceiP04298.

Family & Domainsi

Domains and Repeats

Feature keyPosition(s)LengthDescriptionGraphical viewFeature identifierActions
Domaini516 – 844329mRNA cap 0 methyltransferasePROSITE-ProRule annotationAdd
BLAST

Region

Feature keyPosition(s)LengthDescriptionGraphical viewFeature identifierActions
Regioni1 – 539539Triphosphatase-guanylyltransferaseAdd
BLAST
Regioni549 – 5502S-adenosyl-L-methionine binding
Regioni569 – 5702mRNA cap bindingPROSITE-ProRule annotation
Regioni678 – 6803S-adenosyl-L-methionine binding

Domaini

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.1 Publication

Sequence similaritiesi

In the N-terminal section; belongs to the dsDNA virus mRNA guanylyltransferase family.Curated
In the C-terminal section; belongs to the class I-like SAM-binding methyltransferase superfamily. mRNA cap 0 methyltransferase family.PROSITE-ProRule annotation
Contains 1 mRNA cap 0 methyltransferase domain.PROSITE-ProRule annotation

Family and domain databases

Gene3Di3.40.50.150. 1 hit.
InterProiIPR019602. mRNA_cap_ATPase/GuylTrfase_vir.
IPR004971. mRNA_G-N7_MeTrfase_dom.
IPR029063. SAM-dependent_MTases-like.
[Graphical view]
PfamiPF10640. Pox_ATPase-GT. 1 hit.
PF03291. Pox_MCEL. 1 hit.
[Graphical view]
SUPFAMiSSF53335. SSF53335. 1 hit.
PROSITEiPS51562. RNA_CAP0_MT. 1 hit.
[Graphical view]

Sequencei

Sequence statusi: Complete.

P04298-1 [UniParc]FASTAAdd to Basket

« Hide

        10         20         30         40         50
MDANVVSSST IATYIDALAK NASELEQRST AYEINNELEL VFIKPPLITL
60 70 80 90 100
TNVVNISTIQ ESFIRFTVTN KEGVKIRTKI PLSKVHGLDV KNVQLVDAID
110 120 130 140 150
NIVWEKKSLV TENRLHKECL LRLSTEERHI FLDYKKYGSS IRLELVNLIQ
160 170 180 190 200
AKTKNFTIDF KLKYFLGSGA QSKSSLLHAI NHPKSRPNTS LEIEFTPRDN
210 220 230 240 250
ETVPYDELIK ELTTLSRHIF MASPENVILS PPINAPIKTF MLPKQDIVGL
260 270 280 290 300
DLENLYAVTK TDGIPITIRV TSNGLYCYFT HLGYIIRYPV KRIIDSEVVV
310 320 330 340 350
FGEAVKDKNW TVYLIKLIEP VNAINDRLEE SKYVESKLVD ICDRIVFKSK
360 370 380 390 400
KYEGPFTTTS EVVDMLSTYL PKQPEGVILF YSKGPKSNID FKIKKENTID
410 420 430 440 450
QTANVVFRYM SSEPIIFGES SIFVEYKKFS NDKGFPKEYG SGKIVLYNGV
460 470 480 490 500
NYLNNIYCLE YINTHNEVGI KSVVVPIKFI AEFLVNGEIL KPRIDKTMKY
510 520 530 540 550
INSEDYYGNQ HNIIVEHLRD QSIKIGDIFN EDKLSDVGHQ YANNDKFRLN
560 570 580 590 600
PEVSYFTNKR TRGPLGILSN YVKTLLISMY CSKTFLDDSN KRKVLAIDFG
610 620 630 640 650
NGADLEKYFY GEIALLVATD PDADAIARGN ERYNKLNSGI KTKYYKFDYI
660 670 680 690 700
QETIRSDTFV SSVREVFYFG KFNIIDWQFA IHYSFHPRHY ATVMNNLSEL
710 720 730 740 750
TASGGKVLIT TMDGDKLSKL TDKKTFIIHK NLPSSENYMS VEKIADDRIV
760 770 780 790 800
VYNPSTMSTP MTEYIIKKND IVRVFNEYGF VLVDNVDFAT IIERSKKFIN
810 820 830 840
GASTMEDRPS TRNFFELNRG AIKCEGLDVE DLLSYYVVYV FSKR
Length:844
Mass (Da):96,734
Last modified:March 20, 1987 - v1
Checksum:i8B9FD3DE836FA6E7
GO

Natural variant

Feature keyPosition(s)LengthDescriptionGraphical viewFeature identifierActions
Natural varianti705 – 7051G → D in strain: mutant Dts36.

Sequence databases

Select the link destinations:
EMBL
GenBank
DDBJ
Links Updated
M15058 Genomic DNA. Translation: AAA48253.1.
AY243312 Genomic DNA. Translation: AAO89385.1.
PIRiA03872. QQVZ1.
RefSeqiYP_232988.1. NC_006998.1.

Genome annotation databases

GeneIDi3707562.

Cross-referencesi

Sequence databases

Select the link destinations:
EMBL
GenBank
DDBJ
Links Updated
M15058 Genomic DNA. Translation: AAA48253.1 .
AY243312 Genomic DNA. Translation: AAO89385.1 .
PIRi A03872. QQVZ1.
RefSeqi YP_232988.1. NC_006998.1.

3D structure databases

Select the link destinations:
PDBe
RCSB PDB
PDBj
Links Updated
Entry Method Resolution (Å) Chain Positions PDBsum
2VDW X-ray 2.70 A/C/E/G 545-844 [» ]
4CKB X-ray 2.80 A/D 1-844 [» ]
4CKC X-ray 2.90 A/D 1-844 [» ]
4CKE X-ray 2.90 A/D 1-844 [» ]
ProteinModelPortali P04298.
SMRi P04298. Positions 545-844.
ModBasei Search...
MobiDBi Search...

Protein-protein interaction databases

DIPi DIP-60664N.

Protocols and materials databases

Structural Biology Knowledgebase Search...

Genome annotation databases

GeneIDi 3707562.

Miscellaneous databases

EvolutionaryTracei P04298.

Family and domain databases

Gene3Di 3.40.50.150. 1 hit.
InterProi IPR019602. mRNA_cap_ATPase/GuylTrfase_vir.
IPR004971. mRNA_G-N7_MeTrfase_dom.
IPR029063. SAM-dependent_MTases-like.
[Graphical view ]
Pfami PF10640. Pox_ATPase-GT. 1 hit.
PF03291. Pox_MCEL. 1 hit.
[Graphical view ]
SUPFAMi SSF53335. SSF53335. 1 hit.
PROSITEi PS51562. RNA_CAP0_MT. 1 hit.
[Graphical view ]
ProtoNeti Search...

Publicationsi

« Hide 'large scale' publications
  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.

Entry informationi

Entry nameiMCEL_VACCW
AccessioniPrimary (citable) accession number: P04298
Secondary accession number(s): Q76ZS6
Entry historyi
Integrated into UniProtKB/Swiss-Prot: March 20, 1987
Last sequence update: March 20, 1987
Last modified: October 29, 2014
This is version 82 of the entry and version 1 of the sequence. [Complete history]
Entry statusiReviewed (UniProtKB/Swiss-Prot)
Annotation programViral Protein Annotation Program

Miscellaneousi

Keywords - Technical termi

3D-structure, Complete proteome, Direct protein sequencing, Multifunctional enzyme, Reference proteome

Documents

  1. PDB cross-references
    Index of Protein Data Bank (PDB) cross-references
  2. SIMILARITY comments
    Index of protein domains and families

External Data

Dasty 3