DYR_MOUSE
ID DYR_MOUSE Reviewed; 187 AA.
AC P00375; P70693; Q61485; Q61487; Q61579;
DT 21-JUL-1986, integrated into UniProtKB/Swiss-Prot.
DT 23-JAN-2007, sequence version 3.
DT 03-AUG-2022, entry version 186.
DE RecName: Full=Dihydrofolate reductase;
DE EC=1.5.1.3 {ECO:0000269|PubMed:19748785, ECO:0000269|PubMed:25980602};
GN Name=Dhfr;
OS Mus musculus (Mouse).
OC Eukaryota; Metazoa; Chordata; Craniata; Vertebrata; Euteleostomi; Mammalia;
OC Eutheria; Euarchontoglires; Glires; Rodentia; Myomorpha; Muroidea; Muridae;
OC Murinae; Mus; Mus.
OX NCBI_TaxID=10090;
RN [1]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA].
RX PubMed=6282858; DOI=10.1016/s0021-9258(18)34465-x;
RA Crouse G.F., Simonsen C.C., McEwan R.N., Schimke R.T.;
RT "Structure of amplified normal and variant dihydrofolate reductase genes in
RT mouse sarcoma S180 cells.";
RL J. Biol. Chem. 257:7887-7897(1982).
RN [2]
RP NUCLEOTIDE SEQUENCE [MRNA].
RX PubMed=6573667; DOI=10.1073/pnas.80.9.2495;
RA Simonsen C.C., Levinson A.D.;
RT "Isolation and expression of an altered mouse dihydrofolate reductase
RT cDNA.";
RL Proc. Natl. Acad. Sci. U.S.A. 80:2495-2499(1983).
RN [3]
RP NUCLEOTIDE SEQUENCE [MRNA].
RX PubMed=2263462; DOI=10.1093/nar/18.23.7025;
RA McIvor R.S., Simonsen C.C.;
RT "Isolation and characterization of a variant dihydrofolate reductase cDNA
RT from methotrexate-resistant murine L5178Y cells.";
RL Nucleic Acids Res. 18:7025-7032(1990).
RN [4]
RP NUCLEOTIDE SEQUENCE [LARGE SCALE MRNA].
RC STRAIN=FVB/N; TISSUE=Mammary gland;
RX PubMed=15489334; DOI=10.1101/gr.2596504;
RG The MGC Project Team;
RT "The status, quality, and expansion of the NIH full-length cDNA project:
RT the Mammalian Gene Collection (MGC).";
RL Genome Res. 14:2121-2127(2004).
RN [5]
RP PROTEIN SEQUENCE OF 2-187.
RC TISSUE=Lymphoma;
RX PubMed=762074; DOI=10.1016/s0021-9258(17)37942-5;
RA Stone D., Paterson S.J., Raper J.H., Phillips A.W.;
RT "The amino acid sequence of dihydrofolate reductase from the mouse lymphoma
RT L1210.";
RL J. Biol. Chem. 254:480-488(1979).
RN [6]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA] OF 1-28.
RX PubMed=2982814; DOI=10.1016/s0021-9258(18)89555-2;
RA McGrogan M., Simonsen C.C., Smouse D.T., Farnham P.J., Schimke R.T.;
RT "Heterogeneity at the 5' termini of mouse dihydrofolate reductase mRNAs.
RT Evidence for multiple promoter regions.";
RL J. Biol. Chem. 260:2307-2314(1985).
RN [7]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA / MRNA] OF 1-24; 50-127 AND 154-187.
RX PubMed=6244105; DOI=10.1016/0092-8674(80)90510-3;
RA Nunberg J.H., Kaufman R.J., Chang A.C.Y., Cohen S.N., Schimke R.T.;
RT "Structure and genomic organization of the mouse dihydrofolate reductase
RT gene.";
RL Cell 19:355-364(1980).
RN [8]
RP NUCLEOTIDE SEQUENCE [MRNA] OF 1-24 AND 50-127.
RX PubMed=360074; DOI=10.1038/275617a0;
RA Chang A.C.Y., Nunberg J.H., Kaufman R.J., Erlich H.A., Schimke R.T.,
RA Cohen S.N.;
RT "Phenotypic expression in E. coli of a DNA sequence coding for mouse
RT dihydrofolate reductase.";
RL Nature 275:617-624(1978).
RN [9]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA] OF 163-187.
RX PubMed=6121807; DOI=10.1016/s0021-9258(18)34647-7;
RA Setzer D.R., McGrogan M., Schimke R.T.;
RT "Nucleotide sequence surrounding multiple polyadenylation sites in the
RT mouse dihydrofolate reductase gene.";
RL J. Biol. Chem. 257:5143-5147(1982).
RN [10]
RP IDENTIFICATION BY MASS SPECTROMETRY [LARGE SCALE ANALYSIS].
RC TISSUE=Brain, Heart, Kidney, Liver, Lung, Pancreas, Spleen, and Testis;
RX PubMed=21183079; DOI=10.1016/j.cell.2010.12.001;
RA Huttlin E.L., Jedrychowski M.P., Elias J.E., Goswami T., Rad R.,
RA Beausoleil S.A., Villen J., Haas W., Sowa M.E., Gygi S.P.;
RT "A tissue-specific atlas of mouse protein phosphorylation and expression.";
RL Cell 143:1174-1189(2010).
RN [11]
RP ACETYLATION [LARGE SCALE ANALYSIS] AT LYS-33, SUCCINYLATION [LARGE SCALE
RP ANALYSIS] AT LYS-33, AND IDENTIFICATION BY MASS SPECTROMETRY [LARGE SCALE
RP ANALYSIS].
RC TISSUE=Embryonic fibroblast, and Liver;
RX PubMed=23806337; DOI=10.1016/j.molcel.2013.06.001;
RA Park J., Chen Y., Tishkoff D.X., Peng C., Tan M., Dai L., Xie Z., Zhang Y.,
RA Zwaans B.M., Skinner M.E., Lombard D.B., Zhao Y.;
RT "SIRT5-mediated lysine desuccinylation impacts diverse metabolic
RT pathways.";
RL Mol. Cell 50:919-930(2013).
RN [12]
RP SUBCELLULAR LOCATION, CATALYTIC ACTIVITY, AND FUNCTION.
RX PubMed=25980602; DOI=10.1016/j.febslet.2015.05.017;
RA Hughes L., Carton R., Minguzzi S., McEntee G., Deinum E.E., O'Connell M.J.,
RA Parle-McDermott A.;
RT "An active second dihydrofolate reductase enzyme is not a feature of rat
RT and mouse, but they do have activity in their mitochondria.";
RL FEBS Lett. 589:1855-1862(2015).
RN [13]
RP X-RAY CRYSTALLOGRAPHY (2.50 ANGSTROMS) OF 2-187 IN COMPLEX WITH NADP AND
RP METHOTREXATE, AND CHARACTERIZATION OF METHOTREXATE-RESISTANT VARIANT
RP ARG-23.
RX PubMed=15681865; DOI=10.1107/s0907444904030422;
RA Cody V., Luft J.R., Pangborn W.;
RT "Understanding the role of Leu22 variants in methotrexate resistance:
RT comparison of wild-type and Leu22Arg variant mouse and human dihydrofolate
RT reductase ternary crystal complexes with methotrexate and NADPH.";
RL Acta Crystallogr. D 61:147-155(2005).
RN [14]
RP X-RAY CRYSTALLOGRAPHY (2.00 ANGSTROMS) OF 2-187 IN COMPLEX WITH NADP AND
RP SYNTHETIC INHIBITOR.
RX PubMed=17019704; DOI=10.1002/prot.21131;
RA Cody V., Pace J., Chisum K., Rosowsky A.;
RT "New insights into DHFR interactions: analysis of Pneumocystis carinii and
RT mouse DHFR complexes with NADPH and two highly potent 5-(omega-
RT carboxy(alkyloxy) trimethoprim derivatives reveals conformational
RT correlations with activity and novel parallel ring stacking interactions.";
RL Proteins 65:959-969(2006).
RN [15]
RP X-RAY CRYSTALLOGRAPHY (1.40 ANGSTROMS) IN COMPLEXES WITH NADP AND SYNTHETIC
RP INHIBITOR.
RX PubMed=18703847; DOI=10.1107/s0907444908022348;
RA Cody V., Pace J., Rosowsky A.;
RT "Structural analysis of a holoenzyme complex of mouse dihydrofolate
RT reductase with NADPH and a ternary complex with the potent and selective
RT inhibitor 2,4-diamino-6-(2'-hydroxydibenz[b,f]azepin-5-
RT yl)methylpteridine.";
RL Acta Crystallogr. D 64:977-984(2008).
RN [16]
RP X-RAY CRYSTALLOGRAPHY (1.60 ANGSTROMS) OF 2-187 IN COMPLEXES WITH NADP AND
RP SYNTHETIC INHIBITOR, AND CATALYTIC ACTIVITY.
RX PubMed=19748785; DOI=10.1016/j.bmc.2009.08.044;
RA Gangjee A., Li W., Lin L., Zeng Y., Ihnat M., Warnke L.A., Green D.W.,
RA Cody V., Pace J., Queener S.F.;
RT "Design, synthesis, and X-ray crystal structures of 2,4-diaminofuro[2,3-
RT d]pyrimidines as multireceptor tyrosine kinase and dihydrofolate reductase
RT inhibitors.";
RL Bioorg. Med. Chem. 17:7324-7336(2009).
CC -!- FUNCTION: Key enzyme in folate metabolism. Contributes to the de novo
CC mitochondrial thymidylate biosynthesis pathway (PubMed:25980602).
CC Catalyzes an essential reaction for de novo glycine and purine
CC synthesis, and for DNA precursor synthesis (PubMed:25980602). Binds its
CC own mRNA. {ECO:0000250|UniProtKB:P00374, ECO:0000269|PubMed:25980602}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=(6S)-5,6,7,8-tetrahydrofolate + NADP(+) = 7,8-dihydrofolate +
CC H(+) + NADPH; Xref=Rhea:RHEA:15009, ChEBI:CHEBI:15378,
CC ChEBI:CHEBI:57451, ChEBI:CHEBI:57453, ChEBI:CHEBI:57783,
CC ChEBI:CHEBI:58349; EC=1.5.1.3; Evidence={ECO:0000255|PROSITE-
CC ProRule:PRU00660, ECO:0000269|PubMed:19748785,
CC ECO:0000269|PubMed:25980602};
CC -!- PATHWAY: Cofactor biosynthesis; tetrahydrofolate biosynthesis; 5,6,7,8-
CC tetrahydrofolate from 7,8-dihydrofolate: step 1/1.
CC -!- SUBUNIT: Homodimer. {ECO:0000250|UniProtKB:P00374}.
CC -!- SUBCELLULAR LOCATION: Mitochondrion {ECO:0000269|PubMed:25980602}.
CC Cytoplasm {ECO:0000269|PubMed:25980602}.
CC -!- SIMILARITY: Belongs to the dihydrofolate reductase family.
CC {ECO:0000305}.
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DR EMBL; V00734; CAA24112.1; -; mRNA.
DR EMBL; X56066; CAA39544.1; -; mRNA.
DR EMBL; BC005796; AAH05796.1; -; mRNA.
DR EMBL; M10071; AAA37637.1; -; Genomic_DNA.
DR EMBL; L26316; AAA37523.1; -; mRNA.
DR EMBL; J00387; AAA37638.1; -; Genomic_DNA.
DR EMBL; J00382; AAA37638.1; JOINED; Genomic_DNA.
DR EMBL; J00383; AAA37638.1; JOINED; Genomic_DNA.
DR EMBL; J00384; AAA37638.1; JOINED; Genomic_DNA.
DR EMBL; J00385; AAA37638.1; JOINED; Genomic_DNA.
DR EMBL; J00386; AAA37638.1; JOINED; Genomic_DNA.
DR EMBL; V00731; CAB43539.2; -; mRNA.
DR EMBL; M10722; AAA37524.1; -; mRNA.
DR EMBL; M10811; AAA37525.1; -; mRNA.
DR EMBL; V00733; CAA24111.1; -; Genomic_DNA.
DR CCDS; CCDS26680.1; -.
DR PIR; S13096; RDMSD.
DR RefSeq; NP_034179.1; NM_010049.3.
DR PDB; 1U70; X-ray; 2.50 A; A=2-187.
DR PDB; 2FZJ; X-ray; 2.00 A; A=2-187.
DR PDB; 3D80; X-ray; 1.40 A; A=2-187.
DR PDB; 3D84; X-ray; 1.90 A; X=2-187.
DR PDB; 3K45; X-ray; 1.60 A; A=2-187.
DR PDB; 3K47; X-ray; 2.05 A; A=2-187.
DR PDBsum; 1U70; -.
DR PDBsum; 2FZJ; -.
DR PDBsum; 3D80; -.
DR PDBsum; 3D84; -.
DR PDBsum; 3K45; -.
DR PDBsum; 3K47; -.
DR AlphaFoldDB; P00375; -.
DR SMR; P00375; -.
DR BioGRID; 199218; 11.
DR IntAct; P00375; 1.
DR MINT; P00375; -.
DR STRING; 10090.ENSMUSP00000022218; -.
DR BindingDB; P00375; -.
DR ChEMBL; CHEMBL4564; -.
DR DrugCentral; P00375; -.
DR iPTMnet; P00375; -.
DR PhosphoSitePlus; P00375; -.
DR EPD; P00375; -.
DR jPOST; P00375; -.
DR PaxDb; P00375; -.
DR PeptideAtlas; P00375; -.
DR PRIDE; P00375; -.
DR ProteomicsDB; 277658; -.
DR DNASU; 13361; -.
DR Ensembl; ENSMUST00000022218; ENSMUSP00000022218; ENSMUSG00000021707.
DR GeneID; 13361; -.
DR KEGG; mmu:13361; -.
DR UCSC; uc007rkl.1; mouse.
DR CTD; 1719; -.
DR MGI; MGI:94890; Dhfr.
DR VEuPathDB; HostDB:ENSMUSG00000021707; -.
DR eggNOG; KOG1324; Eukaryota.
DR GeneTree; ENSGT00390000010283; -.
DR HOGENOM; CLU_043966_2_3_1; -.
DR InParanoid; P00375; -.
DR OMA; RDNQLPW; -.
DR OrthoDB; 1489621at2759; -.
DR PhylomeDB; P00375; -.
DR TreeFam; TF317636; -.
DR BRENDA; 1.5.1.3; 3474.
DR Reactome; R-MMU-196757; Metabolism of folate and pterines.
DR SABIO-RK; P00375; -.
DR UniPathway; UPA00077; UER00158.
DR BioGRID-ORCS; 13361; 26 hits in 75 CRISPR screens.
DR ChiTaRS; Dhfr; mouse.
DR EvolutionaryTrace; P00375; -.
DR PRO; PR:P00375; -.
DR Proteomes; UP000000589; Chromosome 13.
DR RNAct; P00375; protein.
DR Bgee; ENSMUSG00000021707; Expressed in optic fissure and 247 other tissues.
DR ExpressionAtlas; P00375; baseline and differential.
DR Genevisible; P00375; MM.
DR GO; GO:0005737; C:cytoplasm; IDA:UniProtKB.
DR GO; GO:0005829; C:cytosol; TAS:Reactome.
DR GO; GO:0005743; C:mitochondrial inner membrane; ISO:MGI.
DR GO; GO:0005759; C:mitochondrial matrix; ISO:MGI.
DR GO; GO:0005739; C:mitochondrion; IDA:UniProtKB.
DR GO; GO:0005634; C:nucleus; IDA:MGI.
DR GO; GO:0004146; F:dihydrofolate reductase activity; IDA:UniProtKB.
DR GO; GO:0051871; F:dihydrofolic acid binding; ISO:MGI.
DR GO; GO:0005542; F:folic acid binding; ISO:MGI.
DR GO; GO:0003729; F:mRNA binding; ISS:UniProtKB.
DR GO; GO:0000900; F:mRNA regulatory element binding translation repressor activity; ISO:MGI.
DR GO; GO:0050661; F:NADP binding; ISO:MGI.
DR GO; GO:0070402; F:NADPH binding; ISO:MGI.
DR GO; GO:1990825; F:sequence-specific mRNA binding; ISO:MGI.
DR GO; GO:0031103; P:axon regeneration; ISO:MGI.
DR GO; GO:0046452; P:dihydrofolate metabolic process; ISO:MGI.
DR GO; GO:0046655; P:folic acid metabolic process; ISO:MGI.
DR GO; GO:0006545; P:glycine biosynthetic process; IEA:InterPro.
DR GO; GO:0051000; P:positive regulation of nitric-oxide synthase activity; IMP:BHF-UCL.
DR GO; GO:2000121; P:regulation of removal of superoxide radicals; IMP:BHF-UCL.
DR GO; GO:0031427; P:response to methotrexate; IEA:UniProtKB-KW.
DR GO; GO:0035094; P:response to nicotine; IEA:Ensembl.
DR GO; GO:0006729; P:tetrahydrobiopterin biosynthetic process; IMP:BHF-UCL.
DR GO; GO:0046654; P:tetrahydrofolate biosynthetic process; ISO:MGI.
DR GO; GO:0035999; P:tetrahydrofolate interconversion; IDA:MGI.
DR GO; GO:0046653; P:tetrahydrofolate metabolic process; ISS:UniProtKB.
DR CDD; cd00209; DHFR; 1.
DR Gene3D; 3.40.430.10; -; 1.
DR InterPro; IPR012259; DHFR.
DR InterPro; IPR024072; DHFR-like_dom_sf.
DR InterPro; IPR017925; DHFR_CS.
DR InterPro; IPR001796; DHFR_dom.
DR PANTHER; PTHR48069; PTHR48069; 1.
DR Pfam; PF00186; DHFR_1; 1.
DR SUPFAM; SSF53597; SSF53597; 1.
DR PROSITE; PS00075; DHFR_1; 1.
DR PROSITE; PS51330; DHFR_2; 1.
PE 1: Evidence at protein level;
KW 3D-structure; Acetylation; Cytoplasm; Direct protein sequencing;
KW Methotrexate resistance; Mitochondrion; NADP; One-carbon metabolism;
KW Oxidoreductase; Reference proteome; RNA-binding.
FT INIT_MET 1
FT /note="Removed"
FT /evidence="ECO:0000269|PubMed:762074"
FT CHAIN 2..187
FT /note="Dihydrofolate reductase"
FT /id="PRO_0000186364"
FT DOMAIN 4..185
FT /note="DHFR"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU00660"
FT BINDING 10
FT /ligand="NADP(+)"
FT /ligand_id="ChEBI:CHEBI:58349"
FT /evidence="ECO:0000269|PubMed:15681865,
FT ECO:0000269|PubMed:17019704"
FT BINDING 16..22
FT /ligand="NADP(+)"
FT /ligand_id="ChEBI:CHEBI:58349"
FT /evidence="ECO:0000269|PubMed:15681865,
FT ECO:0000269|PubMed:17019704"
FT BINDING 31..36
FT /ligand="substrate"
FT /evidence="ECO:0000305|PubMed:15681865"
FT BINDING 55..57
FT /ligand="NADP(+)"
FT /ligand_id="ChEBI:CHEBI:58349"
FT /evidence="ECO:0000269|PubMed:15681865,
FT ECO:0000269|PubMed:17019704"
FT BINDING 65
FT /ligand="substrate"
FT /evidence="ECO:0000305|PubMed:15681865"
FT BINDING 71
FT /ligand="substrate"
FT /evidence="ECO:0000305|PubMed:15681865"
FT BINDING 77..79
FT /ligand="NADP(+)"
FT /ligand_id="ChEBI:CHEBI:58349"
FT /evidence="ECO:0000269|PubMed:15681865,
FT ECO:0000269|PubMed:17019704"
FT BINDING 117..124
FT /ligand="NADP(+)"
FT /ligand_id="ChEBI:CHEBI:58349"
FT /evidence="ECO:0000269|PubMed:15681865,
FT ECO:0000269|PubMed:17019704"
FT MOD_RES 33
FT /note="N6-acetyllysine; alternate"
FT /evidence="ECO:0007744|PubMed:23806337"
FT MOD_RES 33
FT /note="N6-succinyllysine; alternate"
FT /evidence="ECO:0007744|PubMed:23806337"
FT VARIANT 23
FT /note="L -> R (in a form with an abnormally low affinity
FT for methotrexate)"
FT /evidence="ECO:0000269|PubMed:15681865"
FT VARIANT 32
FT /note="F -> W (in L-5178-Y cell line; methotrexate-
FT resistant, requires 2 nucleotide substitutions)"
FT CONFLICT 4
FT /note="P -> A (in Ref. 6; AAA37637)"
FT /evidence="ECO:0000305"
FT CONFLICT 14
FT /note="N -> D (in Ref. 3; CAA39544)"
FT /evidence="ECO:0000305"
FT CONFLICT 123
FT /note="Q -> E (in Ref. 5; AA sequence and 8; AAA37525)"
FT /evidence="ECO:0000305"
FT CONFLICT 124
FT /note="E -> Q (in Ref. 5; AA sequence)"
FT /evidence="ECO:0000305"
FT CONFLICT 128
FT /note="Q -> E (in Ref. 5; AA sequence)"
FT /evidence="ECO:0000305"
FT CONFLICT 174
FT /note="K -> D (in Ref. 5; AA sequence)"
FT /evidence="ECO:0000305"
FT STRAND 4..11
FT /evidence="ECO:0007829|PDB:3D80"
FT STRAND 15..19
FT /evidence="ECO:0007829|PDB:3D80"
FT STRAND 24..26
FT /evidence="ECO:0007829|PDB:3D80"
FT HELIX 29..40
FT /evidence="ECO:0007829|PDB:3D80"
FT STRAND 43..46
FT /evidence="ECO:0007829|PDB:1U70"
FT STRAND 50..54
FT /evidence="ECO:0007829|PDB:3D80"
FT HELIX 55..60
FT /evidence="ECO:0007829|PDB:3D80"
FT HELIX 63..65
FT /evidence="ECO:0007829|PDB:3D80"
FT STRAND 71..76
FT /evidence="ECO:0007829|PDB:3D80"
FT STRAND 88..93
FT /evidence="ECO:0007829|PDB:3D80"
FT HELIX 94..101
FT /evidence="ECO:0007829|PDB:3D80"
FT HELIX 104..109
FT /evidence="ECO:0007829|PDB:3D80"
FT STRAND 113..115
FT /evidence="ECO:0007829|PDB:3D80"
FT HELIX 119..126
FT /evidence="ECO:0007829|PDB:3D80"
FT STRAND 128..130
FT /evidence="ECO:0007829|PDB:3D80"
FT STRAND 132..139
FT /evidence="ECO:0007829|PDB:3D80"
FT STRAND 146..148
FT /evidence="ECO:0007829|PDB:3D80"
FT TURN 154..156
FT /evidence="ECO:0007829|PDB:3D80"
FT STRAND 157..159
FT /evidence="ECO:0007829|PDB:2FZJ"
FT STRAND 171..173
FT /evidence="ECO:0007829|PDB:3D80"
FT STRAND 176..185
FT /evidence="ECO:0007829|PDB:3D80"
SQ SEQUENCE 187 AA; 21606 MW; 47AEF15F879B119C CRC64;
MVRPLNCIVA VSQNMGIGKN GDLPWPPLRN EFKYFQRMTT TSSVEGKQNL VIMGRKTWFS
IPEKNRPLKD RINIVLSREL KEPPRGAHFL AKSLDDALRL IEQPELASKV DMVWIVGGSS
VYQEAMNQPG HLRLFVTRIM QEFESDTFFP EIDLGKYKLL PEYPGVLSEV QEEKGIKYKF
EVYEKKD
