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Gene Review

DHFR  -  dihydrofolate reductase

Homo sapiens

Synonyms: DHFRP1, DYR, Dihydrofolate reductase
Disease relevance of DHFR

Psychiatry related information on DHFR


Disease relevance of DHFR

  • Although the DHFR 19-bp deletion polymorphism was not associated with overall breast cancer risk, we observed a borderline significant additive interaction (P = 0.06) between the DHFR genotype and multivitamin use [8].

High impact information on DHFR

  • The fundamental issues related to this linkage are probed in the context of two enzymes that catalyze hydride transfer, namely dihydrofolate reductase and liver alcohol dehydrogenase [9].
  • Use of multivitamin supplements containing folic acid diminished the adverse effects of dihydrofolate reductase inhibitors, but not that of antiepileptic drugs [10].
  • We have examined DNA repair in the dihydrofolate reductase (DHFR) gene in cultured human cells treated with 4'-hydroxymethyl-4,5',8-trimethylpsoralen (HMT) using a newly developed assay for interstrand DNA cross-linking in defined genomic sequences [11].
  • Selective removal of transcription-blocking DNA damage from the transcribed strand of the mammalian DHFR gene [12].
  • Furthermore, in the 5' flanking region of the human DHFR gene, selective rapid repair occurs in the opposite DNA strand relative to the transcribed strand of the DHFR gene [12].

Chemical compound and disease context of DHFR


Biological context of DHFR


Anatomical context of DHFR

  • The upstream initiating transcripts present in all three cell lines are increased in amount in 6A3 cells as compared with the other cell lines, in about the same proportion as the three identified DHFR mRNAs [21].
  • To investigate the regulated expression of the DHFR gene, we stimulated serum-starved NIH 3T3 cells to synchronously reenter the cell cycle [22].
  • Utilizing a methotrexate-resistant KB cell line designated 1BT, we now report the kinetic basis for altered levels of DHFR RNA observed in FUra-treated cells [23].
  • Phytohemagglutinin-stimulated peripheral blood lymphocytes from 6 patients with increased levels of DHFR showed no evidence of MTX resistance in vitro [24].
  • The CHO cells transformed with the human ERCC-1 gene repaired the active DHFR gene much more efficiently than the non-transcribed sequences, a pattern similar to that seen in wild type CHO cells [25].

Associations of DHFR with chemical compounds

  • The associated increase in DHFR expression resulted in increased resistance to methotrexate but had no effect on other classes of anticancer agents [26].
  • Studies with the recombinant enzyme show that trimethoprim is a very poor inhibitor of P. carinii DHFR and, in fact, is a more potent inhibitor of human DHFR [27].
  • Growth inhibition by MTX and FdUrd was increased and DHFR and TS activities and expression were correspondingly decreased in Rb transfectants of SaOS-2 cells [28].
  • At least six other minor 5' ends have been mapped to nucleotide positions -449 to -480 upstream of the DHFR gene and pertain to approximately 1% of the DHFR-specific polysomal polyadenylic acid-containing RNA [21].
  • Interestingly, pyrimethamine-resistant strains of P. falciparum all have a common point mutation in the DHFR coding sequence (Thr/Ser 108 to Asn), which causes decreased binding of the folate analog [3].
  • Cells with the mutant 3' UTR had a 2-fold increase in DHFR mRNA half-life, expressed higher DHFR mRNA and DHFR protein, and were 4-fold more resistant to methotrexate as compared with WT cells [29].

Physical interactions of DHFR

  • Rb interacts with the family of transcription factors called E2F reducing transcription of genes that contain E2F binding sites in the promoter regions e.g. DHFR [30].
  • The DHFR-TS nucleic acid sequence contains no introns and the single 1563-bp open reading frame encodes a 179 residue N-terminal DHFR domain connected by a 55 amino acid junction peptide to a 287 residue C-terminal TS domain [31].
  • Based on these observations, a new model is proposed whereby DHFR exists in two conformations, one bound to DHFR mRNA and the other bound to NADPH [32].
  • In all recrudescent parasites, however, the presence of Ile 51/Asn 108 dhfr mutations was coupled with the dhps Ala 436 [33].
  • Insertion of a seven amino acid sequence from a structurally equivalent 'beta-blowout' sequence from human DHFR destabilizes E. coli DHFR by 3.6 kcal/mol and decreases catalytic efficiency (kcat/K(m)) 34-fold [34].

Regulatory relationships of DHFR

  • Furthermore, in a cotransfection experiment, expression of human E2F1 stimulated the DHFR promoter 22-fold in serum-starved cells [22].
  • Specifically, TS ligand induced domain-domain communication involving DHFR activation is observed only in the L. major enzyme and, whereas both DHFR activities involve a rate-limiting conformational change, the change occurs at different positions along the kinetic pathway [35].
  • Only Tom20 inhibited the import of a fusion protein of the leader of aldehyde dehydrogenase attached to dihydrofolate reductase [36].
  • By high resolution two-dimensional polyacrylamide gradient electrophoresis, human DHFR was shown to be selectively overproduced in VB2a-100 MTX-resistant cells whereas mouse DHFR protein "spots" present in MTX-sensitive parental hybrid were absent in these cells exhibiting 100 microM MTX resistance [37].
  • Recombinant BHK-21 cells producing hIL-6 under the control of the CMV promoter were contransfected with the ras oncogene and dihydrofolate reductase gene, then selected with 50 nM methotrexate to coamplify the ras oncogene [38].

Other interactions of DHFR

  • Transfectants with increased cyclin D1 expression also had increased DHFR mRNA and protein expression [26].
  • Each reaction resulted in a rapid, time-dependent loss of TS activity and no effect on DHFR activity [39].
  • DHFR/MSH3 amplification in methotrexate-resistant cells alters the hMutSalpha/hMutSbeta ratio and reduces the efficiency of base-base mismatch repair [40].
  • We show that HCMV activates the DHFR promoter and that products of the HCMV major immediate-early gene region mediate the activation of the promoter specifically through the E2F site [41].
  • These data indicate higher FPGS and lower DHFR levels as potential mechanisms contributing to greater MTXPG accumulation and cytotoxicity in B-lineage lymphoblasts [42].

Analytical, diagnostic and therapeutic context of DHFR

  • Dihydrofolate reductase (DHFR) mRNA and protein levels were measured by northern blot and western blot analyses, respectively [26].
  • Most of the recognized elements of secondary structure of other DHFRs, as ascertained by x-ray crystallography, are found in the predicted structure of the DHFR domain of the bifunctional protein [19].
  • Consistent with the recent conclusion that P. carinii is a member of the Fungi, sequence analysis and chromosomal localization show that DHFR is neither physically nor genetically linked to thymidylate synthase [27].
  • We used gel mobility shift assays to search for potential molecular mechanisms for this activation and found an "infection-specific" multimeric complex that bound to the E2F sites in the DHFR and E2 promoters in extracts from HCMV-infected cells but not in extracts from uninfected cells [41].
  • Southern blot analysis with a human DHFR cDNA probe confirmed this increase in the gene copy number, and demonstrated a similar restriction pattern with Eco R1, Hind III, and Pst 1 as seen with a highly amplified human leukemia cell line, K562 [43].



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  12. Selective removal of transcription-blocking DNA damage from the transcribed strand of the mammalian DHFR gene. Mellon, I., Spivak, G., Hanawalt, P.C. Cell (1987) [Pubmed]
  13. Trypanosoma brucei dihydrofolate reductase-thymidylate synthase: gene isolation and expression and characterization of the enzyme. Gamarro, F., Yu, P.L., Zhao, J., Edman, U., Greene, P.J., Santi, D. Mol. Biochem. Parasitol. (1995) [Pubmed]
  14. Protection of hematopoietic stem cells from pemetrexed toxicity by retroviral gene transfer with a mutant dihydrofolate reductase-mutant thymidylate synthase fusion gene. Capiaux, G.M., Budak-Alpdogan, T., Alpdogan, O., Bornmann, W., Takebe, N., Banerjee, D., Maley, F., Bertino, J.R. Cancer Gene Ther. (2004) [Pubmed]
  15. Clinical resistance to antimetabolites. Spears, C.P. Hematol. Oncol. Clin. North Am. (1995) [Pubmed]
  16. Sequence variations in the genes encoding dihydropteroate synthase and dihydrofolate reductase and clinical response to sulfadoxine-pyrimethamine in patients with acute uncomplicated falciparum malaria. Basco, L.K., Tahar, R., Keundjian, A., Ringwald, P. J. Infect. Dis. (2000) [Pubmed]
  17. Chromosomal localization and racial distribution of the polymorphic human dihydrofolate reductase pseudogene (DHFRP1). Anagnou, N.P., Antonarakis, S.E., O'Brien, S.J., Modi, W.S., Nienhuis, A.W. Am. J. Hum. Genet. (1988) [Pubmed]
  18. Coactivator-associated arginine methyltransferase 1 (CARM1) is a positive regulator of the Cyclin E1 gene. El Messaoudi, S., Fabbrizio, E., Rodriguez, C., Chuchana, P., Fauquier, L., Cheng, D., Theillet, C., Vandel, L., Bedford, M.T., Sardet, C. Proc. Natl. Acad. Sci. U.S.A. (2006) [Pubmed]
  19. Bifunctional thymidylate synthase-dihydrofolate reductase from Leishmania tropica: sequence homology with the corresponding monofunctional proteins. Grumont, R., Washtien, W.L., Caput, D., Santi, D.V. Proc. Natl. Acad. Sci. U.S.A. (1986) [Pubmed]
  20. The HIP1 initiator element plays a role in determining the in vitro requirement of the dihydrofolate reductase gene promoter for the C-terminal domain of RNA polymerase II. Buermeyer, A.B., Thompson, N.E., Strasheim, L.A., Burgess, R.R., Farnham, P.J. Mol. Cell. Biol. (1992) [Pubmed]
  21. Discrete human dihydrofolate reductase gene transcripts present in polysomal RNA map with their 5' ends several hundred nucleotides upstream of the main mRNA start site. Masters, J.N., Attardi, G. Mol. Cell. Biol. (1985) [Pubmed]
  22. A protein synthesis-dependent increase in E2F1 mRNA correlates with growth regulation of the dihydrofolate reductase promoter. Slansky, J.E., Li, Y., Kaelin, W.G., Farnham, P.J. Mol. Cell. Biol. (1993) [Pubmed]
  23. 5-Fluorouracil inhibits dihydrofolate reductase precursor mRNA processing and/or nuclear mRNA stability in methotrexate-resistant KB cells. Will, C.L., Dolnick, B.J. J. Biol. Chem. (1989) [Pubmed]
  24. Increase of dihydrofolate reductase in peripheral blood lymphocytes of rheumatoid arthritis patients treated with low-dose oral methotrexate. Rodenhuis, S., Kremer, J.M., Bertino, J.R. Arthritis Rheum. (1987) [Pubmed]
  25. Human repair gene restores normal pattern of preferential DNA repair in repair defective CHO cells. Bohr, V.A., Chu, E.H., van Duin, M., Hanawalt, P.C., Okumoto, D.S. Nucleic Acids Res. (1988) [Pubmed]
  26. Effect of cyclin D1 overexpression on drug sensitivity in a human fibrosarcoma cell line. Hochhauser, D., Schnieders, B., Ercikan-Abali, E., Gorlick, R., Muise-Helmericks, R., Li, W.W., Fan, J., Banerjee, D., Bertino, J.R. J. Natl. Cancer Inst. (1996) [Pubmed]
  27. Isolation and expression of the Pneumocystis carinii dihydrofolate reductase gene. Edman, J.C., Edman, U., Cao, M., Lundgren, B., Kovacs, J.A., Santi, D.V. Proc. Natl. Acad. Sci. U.S.A. (1989) [Pubmed]
  28. Lack of functional retinoblastoma protein mediates increased resistance to antimetabolites in human sarcoma cell lines. Li, W., Fan, J., Hochhauser, D., Banerjee, D., Zielinski, Z., Almasan, A., Yin, Y., Kelly, R., Wahl, G.M., Bertino, J.R. Proc. Natl. Acad. Sci. U.S.A. (1995) [Pubmed]
  29. A miR-24 microRNA binding-site polymorphism in dihydrofolate reductase gene leads to methotrexate resistance. Mishra, P.J., Humeniuk, R., Mishra, P.J., Longo-Sorbello, G.S., Banerjee, D., Bertino, J.R. Proc. Natl. Acad. Sci. U.S.A. (2007) [Pubmed]
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  32. Identification of amino acids required for the functional up-regulation of human dihydrofolate reductase protein in response to antifolate Treatment. Skacel, N., Menon, L.G., Mishra, P.J., Peters, R., Banerjee, D., Bertino, J.R., Abali, E.E. J. Biol. Chem. (2005) [Pubmed]
  33. Pyrimethamine/sulfadoxine combination in the treatment of uncomplicated falciparum malaria: relation between dihydropteroate synthase/dihydrofolate reductase genotypes, sulfadoxine plasma levels, and treatment outcome. Khalil, I., Alifrangis, M., Rønn, A.M., Gabar, H.A., Jelinek, T., Satti, G.M., Bygbjerg, I.C. Am. J. Trop. Med. Hyg. (2002) [Pubmed]
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  40. DHFR/MSH3 amplification in methotrexate-resistant cells alters the hMutSalpha/hMutSbeta ratio and reduces the efficiency of base-base mismatch repair. Drummond, J.T., Genschel, J., Wolf, E., Modrich, P. Proc. Natl. Acad. Sci. U.S.A. (1997) [Pubmed]
  41. E2F mediates dihydrofolate reductase promoter activation and multiprotein complex formation in human cytomegalovirus infection. Wade, M., Kowalik, T.F., Mudryj, M., Huang, E.S., Azizkhan, J.C. Mol. Cell. Biol. (1992) [Pubmed]
  42. Differences in folylpolyglutamate synthetase and dihydrofolate reductase expression in human B-lineage versus T-lineage leukemic lymphoblasts: mechanisms for lineage differences in methotrexate polyglutamylation and cytotoxicity. Galpin, A.J., Schuetz, J.D., Masson, E., Yanishevski, Y., Synold, T.W., Barredo, J.C., Pui, C.H., Relling, M.V., Evans, W.E. Mol. Pharmacol. (1997) [Pubmed]
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