5NT3B_DROME
ID 5NT3B_DROME Reviewed; 319 AA.
AC Q9W197;
DT 03-APR-2013, integrated into UniProtKB/Swiss-Prot.
DT 01-MAY-2000, sequence version 1.
DT 03-AUG-2022, entry version 139.
DE RecName: Full=7-methylguanosine phosphate-specific 5'-nucleotidase;
DE Short=7-methylguanosine nucleotidase;
DE EC=3.1.3.91 {ECO:0000269|PubMed:23223233, ECO:0000269|PubMed:24603684};
DE AltName: Full=Cytosolic 5'-nucleotidase IIIB {ECO:0000312|FlyBase:FBgn0034988};
DE EC=3.1.3.5 {ECO:0000269|PubMed:23223233, ECO:0000269|PubMed:24603684};
DE AltName: Full=N(7)-methylguanylate 5'-phosphatase;
GN Name=cN-IIIB {ECO:0000312|FlyBase:FBgn0034988};
GN ORFNames=CG3362 {ECO:0000312|FlyBase:FBgn0034988};
OS Drosophila melanogaster (Fruit fly).
OC Eukaryota; Metazoa; Ecdysozoa; Arthropoda; Hexapoda; Insecta; Pterygota;
OC Neoptera; Endopterygota; Diptera; Brachycera; Muscomorpha; Ephydroidea;
OC Drosophilidae; Drosophila; Sophophora.
OX NCBI_TaxID=7227;
RN [1]
RP NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
RC STRAIN=Berkeley;
RX PubMed=10731132; DOI=10.1126/science.287.5461.2185;
RA Adams M.D., Celniker S.E., Holt R.A., Evans C.A., Gocayne J.D.,
RA Amanatides P.G., Scherer S.E., Li P.W., Hoskins R.A., Galle R.F.,
RA George R.A., Lewis S.E., Richards S., Ashburner M., Henderson S.N.,
RA Sutton G.G., Wortman J.R., Yandell M.D., Zhang Q., Chen L.X., Brandon R.C.,
RA Rogers Y.-H.C., Blazej R.G., Champe M., Pfeiffer B.D., Wan K.H., Doyle C.,
RA Baxter E.G., Helt G., Nelson C.R., Miklos G.L.G., Abril J.F., Agbayani A.,
RA An H.-J., Andrews-Pfannkoch C., Baldwin D., Ballew R.M., Basu A.,
RA Baxendale J., Bayraktaroglu L., Beasley E.M., Beeson K.Y., Benos P.V.,
RA Berman B.P., Bhandari D., Bolshakov S., Borkova D., Botchan M.R., Bouck J.,
RA Brokstein P., Brottier P., Burtis K.C., Busam D.A., Butler H., Cadieu E.,
RA Center A., Chandra I., Cherry J.M., Cawley S., Dahlke C., Davenport L.B.,
RA Davies P., de Pablos B., Delcher A., Deng Z., Mays A.D., Dew I.,
RA Dietz S.M., Dodson K., Doup L.E., Downes M., Dugan-Rocha S., Dunkov B.C.,
RA Dunn P., Durbin K.J., Evangelista C.C., Ferraz C., Ferriera S.,
RA Fleischmann W., Fosler C., Gabrielian A.E., Garg N.S., Gelbart W.M.,
RA Glasser K., Glodek A., Gong F., Gorrell J.H., Gu Z., Guan P., Harris M.,
RA Harris N.L., Harvey D.A., Heiman T.J., Hernandez J.R., Houck J., Hostin D.,
RA Houston K.A., Howland T.J., Wei M.-H., Ibegwam C., Jalali M., Kalush F.,
RA Karpen G.H., Ke Z., Kennison J.A., Ketchum K.A., Kimmel B.E., Kodira C.D.,
RA Kraft C.L., Kravitz S., Kulp D., Lai Z., Lasko P., Lei Y., Levitsky A.A.,
RA Li J.H., Li Z., Liang Y., Lin X., Liu X., Mattei B., McIntosh T.C.,
RA McLeod M.P., McPherson D., Merkulov G., Milshina N.V., Mobarry C.,
RA Morris J., Moshrefi A., Mount S.M., Moy M., Murphy B., Murphy L.,
RA Muzny D.M., Nelson D.L., Nelson D.R., Nelson K.A., Nixon K., Nusskern D.R.,
RA Pacleb J.M., Palazzolo M., Pittman G.S., Pan S., Pollard J., Puri V.,
RA Reese M.G., Reinert K., Remington K., Saunders R.D.C., Scheeler F.,
RA Shen H., Shue B.C., Siden-Kiamos I., Simpson M., Skupski M.P., Smith T.J.,
RA Spier E., Spradling A.C., Stapleton M., Strong R., Sun E., Svirskas R.,
RA Tector C., Turner R., Venter E., Wang A.H., Wang X., Wang Z.-Y.,
RA Wassarman D.A., Weinstock G.M., Weissenbach J., Williams S.M., Woodage T.,
RA Worley K.C., Wu D., Yang S., Yao Q.A., Ye J., Yeh R.-F., Zaveri J.S.,
RA Zhan M., Zhang G., Zhao Q., Zheng L., Zheng X.H., Zhong F.N., Zhong W.,
RA Zhou X., Zhu S.C., Zhu X., Smith H.O., Gibbs R.A., Myers E.W., Rubin G.M.,
RA Venter J.C.;
RT "The genome sequence of Drosophila melanogaster.";
RL Science 287:2185-2195(2000).
RN [2]
RP GENOME REANNOTATION.
RC STRAIN=Berkeley;
RX PubMed=12537572; DOI=10.1186/gb-2002-3-12-research0083;
RA Misra S., Crosby M.A., Mungall C.J., Matthews B.B., Campbell K.S.,
RA Hradecky P., Huang Y., Kaminker J.S., Millburn G.H., Prochnik S.E.,
RA Smith C.D., Tupy J.L., Whitfield E.J., Bayraktaroglu L., Berman B.P.,
RA Bettencourt B.R., Celniker S.E., de Grey A.D.N.J., Drysdale R.A.,
RA Harris N.L., Richter J., Russo S., Schroeder A.J., Shu S.Q., Stapleton M.,
RA Yamada C., Ashburner M., Gelbart W.M., Rubin G.M., Lewis S.E.;
RT "Annotation of the Drosophila melanogaster euchromatic genome: a systematic
RT review.";
RL Genome Biol. 3:RESEARCH0083.1-RESEARCH0083.22(2002).
RN [3]
RP NUCLEOTIDE SEQUENCE [LARGE SCALE MRNA].
RC STRAIN=Berkeley; TISSUE=Embryo;
RX PubMed=12537569; DOI=10.1186/gb-2002-3-12-research0080;
RA Stapleton M., Carlson J.W., Brokstein P., Yu C., Champe M., George R.A.,
RA Guarin H., Kronmiller B., Pacleb J.M., Park S., Wan K.H., Rubin G.M.,
RA Celniker S.E.;
RT "A Drosophila full-length cDNA resource.";
RL Genome Biol. 3:RESEARCH0080.1-RESEARCH0080.8(2002).
RN [4]
RP FUNCTION, CATALYTIC ACTIVITY, BIOPHYSICOCHEMICAL PROPERTIES, ACTIVITY
RP REGULATION, AND SUBUNIT.
RX PubMed=23223233; DOI=10.1074/jbc.m112.426700;
RA Buschmann J., Moritz B., Jeske M., Lilie H., Schierhorn A., Wahle E.;
RT "Identification of Drosophila and Human 7-Methyl GMP-specific
RT Nucleotidases.";
RL J. Biol. Chem. 288:2441-2451(2013).
RN [5]
RP X-RAY CRYSTALLOGRAPHY (1.65 ANGSTROMS) IN COMPLEX WITH REACTION PRODUCTS,
RP FUNCTION, CATALYTIC ACTIVITY, COFACTOR, BIOPHYSICOCHEMICAL PROPERTIES,
RP ACTIVE SITE, METAL BINDING, AND MUTAGENESIS OF PHE-75; GLY-76; TRP-121 AND
RP 75-PHE--GLY-76.
RX PubMed=24603684; DOI=10.1371/journal.pone.0090915;
RA Monecke T., Buschmann J., Neumann P., Wahle E., Ficner R.;
RT "Crystal structures of the novel cytosolic 5'-nucleotidase IIIB explain its
RT preference for m7GMP.";
RL PLoS ONE 9:90915-90915(2014).
CC -!- FUNCTION: Specifically hydrolyzes 7-methylguanosine monophosphate
CC (m(7)GMP) to 7-methylguanosine and inorganic phosphate
CC (PubMed:23223233, PubMed:24603684). Also able to mediate hydrolysis of
CC diphosphate (m(7)GDP) to 7-methylguanosine and 2 inorganic phosphate
CC with lower activity (PubMed:23223233). The specific activity for
CC m(7)GMP may protect cells against undesired salvage of m(7)GMP and its
CC incorporation into nucleic acids (PubMed:23223233). Also has weak
CC activity for CMP (PubMed:23223233, PubMed:24603684). UMP and purine
CC nucleotides are poor substrates (PubMed:23223233, PubMed:24603684).
CC {ECO:0000269|PubMed:23223233, ECO:0000269|PubMed:24603684}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=H2O + N(7)-methyl-GMP = N(7)-methylguanosine + phosphate;
CC Xref=Rhea:RHEA:37107, ChEBI:CHEBI:15377, ChEBI:CHEBI:20794,
CC ChEBI:CHEBI:43474, ChEBI:CHEBI:58285; EC=3.1.3.91;
CC Evidence={ECO:0000269|PubMed:23223233, ECO:0000269|PubMed:24603684};
CC -!- CATALYTIC ACTIVITY:
CC Reaction=CMP + H2O = cytidine + phosphate; Xref=Rhea:RHEA:29367,
CC ChEBI:CHEBI:15377, ChEBI:CHEBI:17562, ChEBI:CHEBI:43474,
CC ChEBI:CHEBI:60377; EC=3.1.3.91;
CC Evidence={ECO:0000269|PubMed:23223233, ECO:0000269|PubMed:24603684};
CC -!- CATALYTIC ACTIVITY:
CC Reaction=a ribonucleoside 5'-phosphate + H2O = a ribonucleoside +
CC phosphate; Xref=Rhea:RHEA:12484, ChEBI:CHEBI:15377,
CC ChEBI:CHEBI:18254, ChEBI:CHEBI:43474, ChEBI:CHEBI:58043; EC=3.1.3.5;
CC Evidence={ECO:0000269|PubMed:23223233, ECO:0000269|PubMed:24603684};
CC -!- COFACTOR:
CC Name=Mg(2+); Xref=ChEBI:CHEBI:18420;
CC Evidence={ECO:0000269|PubMed:24603684};
CC -!- ACTIVITY REGULATION: Inhibited by high levels of AMP.
CC {ECO:0000269|PubMed:23223233}.
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=13 uM for m(7)GMP (at 25 degrees Celsius)
CC {ECO:0000269|PubMed:23223233};
CC KM=12 uM for m(7)GMP (at 25 degrees Celsius)
CC {ECO:0000269|PubMed:24603684};
CC KM=48 uM for CMP (at 25 degrees Celsius)
CC {ECO:0000269|PubMed:23223233, ECO:0000269|PubMed:24603684};
CC KM=102 uM for GMP (at 25 degrees Celsius)
CC {ECO:0000269|PubMed:23223233};
CC KM=32 uM for AMP (at 25 degrees Celsius)
CC {ECO:0000269|PubMed:23223233};
CC KM=91 uM for UMP (at 25 degrees Celsius)
CC {ECO:0000269|PubMed:23223233};
CC Vmax=11 umol/min/mg enzyme with m(7)GMP as substrate
CC {ECO:0000269|PubMed:23223233};
CC Vmax=21 umol/min/mg enzyme with CMP as substrate
CC {ECO:0000269|PubMed:23223233};
CC Vmax=1.5 umol/min/mg enzyme with GMP as substrate
CC {ECO:0000269|PubMed:23223233};
CC Vmax=0.49 umol/min/mg enzyme with AMP as substrate
CC {ECO:0000269|PubMed:23223233};
CC Vmax=4.0 umol/min/mg enzyme with UMP as substrate
CC {ECO:0000269|PubMed:23223233};
CC Note=kcat is 6.3 sec(-1) with m(7)GMP. kcat is 12 sec(-1) with CMP.
CC kcat is 0.88 sec(-1) with GMP. kcat is 0.28 sec(-1) with AMP. kcat is
CC 2.3 sec(-1) with UMP.;
CC pH dependence:
CC Optimum pH is 7.5. {ECO:0000269|PubMed:23223233};
CC -!- SUBUNIT: Monomer. {ECO:0000269|PubMed:23223233}.
CC -!- SIMILARITY: Belongs to the pyrimidine 5'-nucleotidase family.
CC {ECO:0000305}.
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DR EMBL; AE013599; AAF47180.1; -; Genomic_DNA.
DR EMBL; AY061358; AAL28906.1; -; mRNA.
DR RefSeq; NP_611895.1; NM_138051.4.
DR PDB; 4NV0; X-ray; 1.65 A; A/B=1-319.
DR PDB; 4NWI; X-ray; 2.05 A; A/B=1-319.
DR PDBsum; 4NV0; -.
DR PDBsum; 4NWI; -.
DR AlphaFoldDB; Q9W197; -.
DR SMR; Q9W197; -.
DR IntAct; Q9W197; 1.
DR STRING; 7227.FBpp0072168; -.
DR PaxDb; Q9W197; -.
DR PRIDE; Q9W197; -.
DR DNASU; 37875; -.
DR EnsemblMetazoa; FBtr0072261; FBpp0072168; FBgn0034988.
DR GeneID; 37875; -.
DR KEGG; dme:Dmel_CG3362; -.
DR UCSC; CG3362-RA; d. melanogaster.
DR CTD; 37875; -.
DR FlyBase; FBgn0034988; cN-IIIB.
DR VEuPathDB; VectorBase:FBgn0034988; -.
DR eggNOG; KOG3128; Eukaryota.
DR GeneTree; ENSGT00390000012959; -.
DR HOGENOM; CLU_048584_0_2_1; -.
DR InParanoid; Q9W197; -.
DR OMA; GPERMQI; -.
DR OrthoDB; 1171042at2759; -.
DR PhylomeDB; Q9W197; -.
DR BRENDA; 3.1.3.91; 1994.
DR Reactome; R-DME-429958; mRNA decay by 3' to 5' exoribonuclease.
DR Reactome; R-DME-73621; Pyrimidine catabolism.
DR SABIO-RK; Q9W197; -.
DR BioGRID-ORCS; 37875; 0 hits in 3 CRISPR screens.
DR GenomeRNAi; 37875; -.
DR PRO; PR:Q9W197; -.
DR Proteomes; UP000000803; Chromosome 2R.
DR Bgee; FBgn0034988; Expressed in adult midgut (Drosophila) and 25 other tissues.
DR Genevisible; Q9W197; DM.
DR GO; GO:0005737; C:cytoplasm; IBA:GO_Central.
DR GO; GO:0005829; C:cytosol; IDA:FlyBase.
DR GO; GO:0008253; F:5'-nucleotidase activity; IDA:FlyBase.
DR GO; GO:0000287; F:magnesium ion binding; IEA:InterPro.
DR GO; GO:0008252; F:nucleotidase activity; IDA:FlyBase.
DR GO; GO:0000166; F:nucleotide binding; IEA:UniProtKB-KW.
DR GO; GO:0106411; F:XMP 5'-nucleosidase activity; IEA:UniProtKB-EC.
DR GO; GO:0016311; P:dephosphorylation; IDA:FlyBase.
DR GO; GO:1901069; P:guanosine-containing compound catabolic process; IDA:FlyBase.
DR GO; GO:0009117; P:nucleotide metabolic process; IEA:UniProtKB-KW.
DR GO; GO:0009158; P:ribonucleoside monophosphate catabolic process; IDA:FlyBase.
DR CDD; cd07504; HAD_5NT; 1.
DR Gene3D; 3.40.50.1000; -; 1.
DR InterPro; IPR036412; HAD-like_sf.
DR InterPro; IPR023214; HAD_sf.
DR InterPro; IPR006434; Pyrimidine_nucleotidase_eu.
DR Pfam; PF05822; UMPH-1; 1.
DR SFLD; SFLDG01128; C1.4:_5'-Nucleotidase_Like; 1.
DR SUPFAM; SSF56784; SSF56784; 1.
DR TIGRFAMs; TIGR01544; HAD-SF-IE; 1.
PE 1: Evidence at protein level;
KW 3D-structure; Hydrolase; Magnesium; Metal-binding; Nucleotide metabolism;
KW Nucleotide-binding; Reference proteome.
FT CHAIN 1..319
FT /note="7-methylguanosine phosphate-specific 5'-
FT nucleotidase"
FT /id="PRO_0000421994"
FT ACT_SITE 55
FT /note="Nucleophile"
FT /evidence="ECO:0000269|PubMed:24603684"
FT ACT_SITE 57
FT /note="Proton donor"
FT /evidence="ECO:0000269|PubMed:24603684"
FT BINDING 55
FT /ligand="Mg(2+)"
FT /ligand_id="ChEBI:CHEBI:18420"
FT /evidence="ECO:0000269|PubMed:24603684"
FT BINDING 57
FT /ligand="Mg(2+)"
FT /ligand_id="ChEBI:CHEBI:18420"
FT /evidence="ECO:0000269|PubMed:24603684"
FT BINDING 103
FT /ligand="CMP"
FT /ligand_id="ChEBI:CHEBI:60377"
FT /evidence="ECO:0000269|PubMed:24603684"
FT BINDING 103
FT /ligand="N(7)-methyl-GMP"
FT /ligand_id="ChEBI:CHEBI:58285"
FT /evidence="ECO:0000269|PubMed:24603684"
FT BINDING 124
FT /ligand="N(7)-methyl-GMP"
FT /ligand_id="ChEBI:CHEBI:58285"
FT /evidence="ECO:0000269|PubMed:24603684"
FT BINDING 171..172
FT /ligand="substrate"
FT /evidence="ECO:0000250|UniProtKB:Q9H0P0"
FT BINDING 245
FT /ligand="Mg(2+)"
FT /ligand_id="ChEBI:CHEBI:18420"
FT /evidence="ECO:0000269|PubMed:24603684"
FT MUTAGEN 75..76
FT /note="FG->HN: Increases KM 7-fold for CMP; when associated
FT with TRP-121."
FT /evidence="ECO:0000269|PubMed:24603684"
FT MUTAGEN 75
FT /note="F->H: Increases KM 3-fold for m(7)GMP and increases
FT KM 10-fold for CMP. Decreases KM 6-fold for m(7)GMP and
FT increases KM 2-fold for CMP; when associated with TRP-121."
FT /evidence="ECO:0000269|PubMed:24603684"
FT MUTAGEN 76
FT /note="G->N: Decreases KM 6-fold for m(7)GMP. Decreases KM
FT 1.5-fold for m(7)GMP and increases KM 1.5-fold for CMP;
FT when associated with TRP-121."
FT /evidence="ECO:0000269|PubMed:24603684"
FT MUTAGEN 121
FT /note="W->Y: Considerable increase (12-fold) in KM for
FT m(7)GMP and decreases KM 3-fold for CMP. Decreases KM 6-
FT fold for m(7)GMP and increases KM 2-fold for CMP; when
FT associated with HIS-75. Decreases KM 1.5-fold for m(7)GMP
FT and increases KM 1.5-fold for CMP; when associated with
FT ASN-76. Increases KM 7-fold for CMP; when associated with
FT 75-HIS--ASN-76."
FT HELIX 16..18
FT /evidence="ECO:0007829|PDB:4NV0"
FT HELIX 20..23
FT /evidence="ECO:0007829|PDB:4NV0"
FT HELIX 32..45
FT /evidence="ECO:0007829|PDB:4NV0"
FT HELIX 47..49
FT /evidence="ECO:0007829|PDB:4NV0"
FT STRAND 50..54
FT /evidence="ECO:0007829|PDB:4NV0"
FT TURN 57..59
FT /evidence="ECO:0007829|PDB:4NV0"
FT STRAND 66..68
FT /evidence="ECO:0007829|PDB:4NWI"
FT HELIX 74..79
FT /evidence="ECO:0007829|PDB:4NV0"
FT STRAND 82..84
FT /evidence="ECO:0007829|PDB:4NWI"
FT HELIX 88..103
FT /evidence="ECO:0007829|PDB:4NV0"
FT STRAND 106..108
FT /evidence="ECO:0007829|PDB:4NV0"
FT HELIX 110..127
FT /evidence="ECO:0007829|PDB:4NV0"
FT STRAND 128..131
FT /evidence="ECO:0007829|PDB:4NV0"
FT HELIX 135..142
FT /evidence="ECO:0007829|PDB:4NV0"
FT HELIX 143..148
FT /evidence="ECO:0007829|PDB:4NV0"
FT HELIX 153..163
FT /evidence="ECO:0007829|PDB:4NV0"
FT STRAND 167..174
FT /evidence="ECO:0007829|PDB:4NV0"
FT HELIX 175..184
FT /evidence="ECO:0007829|PDB:4NV0"
FT STRAND 192..197
FT /evidence="ECO:0007829|PDB:4NV0"
FT STRAND 199..202
FT /evidence="ECO:0007829|PDB:4NV0"
FT STRAND 205..209
FT /evidence="ECO:0007829|PDB:4NV0"
FT TURN 221..223
FT /evidence="ECO:0007829|PDB:4NV0"
FT HELIX 231..234
FT /evidence="ECO:0007829|PDB:4NV0"
FT STRAND 239..246
FT /evidence="ECO:0007829|PDB:4NV0"
FT HELIX 247..249
FT /evidence="ECO:0007829|PDB:4NV0"
FT TURN 250..255
FT /evidence="ECO:0007829|PDB:4NV0"
FT STRAND 261..268
FT /evidence="ECO:0007829|PDB:4NV0"
FT HELIX 272..282
FT /evidence="ECO:0007829|PDB:4NV0"
FT STRAND 284..289
FT /evidence="ECO:0007829|PDB:4NV0"
FT HELIX 294..310
FT /evidence="ECO:0007829|PDB:4NV0"
SQ SEQUENCE 319 AA; 36294 MW; 71919CE1CBF34EB5 CRC64;
MGFDEKREPT GGRLRLQDIP ALTQDHCRMR DPAEVERIIN EFVIGGPERM QIVSDFDYTI
TKQRTEDGGA VPSSFGIFNA CQSLPENFKA ETDKLYHKYR PIEIDPHMPI AEKVQYMIEW
WTKSGELTSG FPFDQSEIDQ IASKYTHALR DRTHEFFADL QRLGIPTLVF SAGLGNSVVS
VLRQANVLHP NVKVVSNFLQ FRDGLLDGFQ QPMIHTFNKN ETVLNETSEY YDLVHTRDHI
IVMGDSIGDA DMASGVPASS HIMKIGFLFD HVEANMKKYM DTFDIVLVDD QTMDVPRTLL
SLIEKQHKLN LEAPKQSSL
