Affinage

DHFR

Dihydrofolate reductase · UniProt P00374

Length
187 aa
Mass
21.5 kDa
Annotated
2026-06-09
100 papers in source corpus 15 papers cited in narrative 15 extracted findings
Cross-family judge vs UniProt: tie faithfulness: 8/8 claims corpus-supported (100%)

Mechanistic narrative

Synthesis pass · prose summary of the discoveries below

DHFR is an NADPH-dependent oxidoreductase central to one-carbon metabolism, catalyzing reduction of dihydrofolate to tetrahydrofolate to maintain cellular folate pools required for purine/pyrimidine synthesis and downstream physiology (PMID:21310277). Loss of function has direct clinical consequences: a homozygous p.Asp153Val mutation severely reduces DHFR enzymatic activity and protein (but not mRNA) levels, causing megaloblastic anemia and cerebral folate deficiency, and demonstrating that DHFR is needed to maintain CSF and red-cell folate even when plasma folate is adequate (PMID:21310277). In the vascular endothelium DHFR regenerates tetrahydrobiopterin (H4B) from folic acid-derived substrate, and this activity is required for eNOS recoupling and nitric oxide bioavailability; DHFR knockdown or methotrexate abolishes folic-acid-mediated restoration of NO, and TGFβ-driven DHFR downregulation uncouples eNOS to promote aortic aneurysm in a feed-forward loop reversible by folic acid (PMID:19660467, PMID:33126053). DHFR activity also supports self-renewal and tumorigenicity of brain tumor-initiating cells through one-carbon metabolism (PMID:33545223). At the catalytic level, mechanistic studies in the bacterial ortholog establish that hydride transfer is independent of overall protein mass but depends on appropriately tuned, residue-specific coupled motions—a conserved active-site residue (Asp122) is required for optimal catalysis, illustrating a Goldilocks principle of enzyme flexibility, and ligand binding reorganizes the residue interaction network around the active site (PMID:26652185, PMID:30040418, PMID:17571919). DHFR transcription is cell-cycle regulated: Sp1 is constitutively bound to upstream promoter sites while a cell-cycle-regulated E2F species activates transcription at G1-to-S, and promoter repression upon quiescence is enforced by cooperative Sp1-pRb-HDAC1 and p130-E2F4-DP1 complexes (PMID:11158299, PMID:8552092). The amplified DHFR locus has also served as a model for transcription-coupled DNA repair, where pyrimidine dimer removal is preferential in the active gene (PMID:3838150). Pharmacologically, DHFR can be eliminated by methotrexate-based PROTACs, whose degradation phenotype differs from enzymatic inhibition (PMID:37875111).

Mechanistic history

Synthesis pass · year-by-year structured walk · 14 steps
  1. 1985 High

    Established that DNA repair efficiency is coupled to transcriptional activity using the amplified DHFR locus as a tractable model, answering whether active genes are repaired preferentially.

    Evidence UV-endonuclease quantitation of pyrimidine dimers in defined DHFR restriction fragments in amplified CHO cells

    PMID:3838150

    Open questions at the time
    • Does not identify the repair factors mediating preferential repair
    • Specific to UV pyrimidine dimers, not generalizable to all lesion types
  2. 1991 High

    Resolved whether DHFR downregulation in quiescent/differentiated cells is transcriptional or post-transcriptional, showing it is fully accounted for by reduced mRNA with unchanged translational efficiency.

    Evidence Metabolic labeling, Northern blot, and polysome fractionation in differentiated myoblasts with amplified DHFR

    PMID:2046674

    Open questions at the time
    • Does not define the transcription factors driving the mRNA decrease
    • Limited to myogenic differentiation context
  3. 1993 Medium

    Tested whether transcription-coupled repair is lesion-general, showing N-methylpurine base-excision repair is NOT preferential in the active DHFR gene, delimiting the scope of transcription-coupled repair.

    Evidence Alkaline hydrolysis quantitation of AP sites at DHFR, flanking, and mitochondrial DNA in CHO cells

    PMID:8222061

    Open questions at the time
    • Negative result; single lab
    • Does not address other base lesions or repair pathways
  4. 1996 High

    Defined the cell-cycle architecture of the DHFR promoter, distinguishing constitutive Sp1 binding from a cell-cycle-regulated E2F species that activates transcription at G1-to-S.

    Evidence High-resolution genomic DNase I footprinting and gel mobility shift with sequence-specific probes across the cell cycle

    PMID:8552092

    Open questions at the time
    • Does not identify the specific E2F family member by isoform
    • Footprinting in CHO amplified locus may not capture single-copy regulation
  5. 1999 Medium

    Asked whether transcription resumes only after lesion repair, showing RNA polymerase II can bypass UV lesions in DHFR before they are removed.

    Evidence In vivo RNA labeling and nuclear run-on at three DHFR positions in UV-irradiated CHO cells

    PMID:10190552

    Open questions at the time
    • Compared to published photolesion kinetics from a separate study rather than matched controls
    • Mechanism of polymerase bypass not defined
  6. 2000 Medium

    Characterized RIP60 as a zinc-finger protein binding the DHFR replication origin oribeta, looping DNA via defined zinc-finger hands, linking the locus to origin function.

    Evidence One-hybrid cloning, footprinting, gel shift, AFM, and origin-dependent replication assay

    PMID:10606657

    Open questions at the time
    • RIP60 replication enhancer activity is weak; physiological role at oribeta unproven
    • Does not establish requirement for DHFR origin firing in vivo
  7. 2001 High

    Mechanistically dissected DHFR promoter repression during cell-cycle withdrawal into two cooperating complexes (Sp1-pRb-HDAC1 and p130-E2F4-DP1), explaining how histone deacetylation and pocket proteins coordinate repression.

    Evidence Reciprocal Co-IP, TSA treatment, reporter assays, and GAL4 recruitment in CHOC400 cells

    PMID:11158299

    Open questions at the time
    • Does not map chromatin changes at the endogenous single-copy promoter genome-wide
    • Quantitative contribution of each complex to repression unresolved
  8. 2007 Medium

    Addressed how the active site coordinates catalysis at the level of protein structure, showing ligand binding reorganizes the residue interaction network through the cofactor and that active-site residues have highest network centrality.

    Evidence Residue interaction network analysis and molecular dynamics of native and 19 circularly permuted DHFRs

    PMID:17571919

    Open questions at the time
    • Computational only; no in vitro validation of permuted variants
    • Network predictions not tied to measured catalytic parameters
  9. 2009 High

    Established DHFR as the required intermediate for folic-acid-mediated endothelial protection, regenerating H4B to recouple eNOS and restore NO.

    Evidence HPLC-fluorescence DHFR activity assay, siRNA knockdown, methotrexate inhibition, and NO/superoxide measurements in endothelial cells and Ang II mice

    PMID:19660467

    Open questions at the time
    • Does not define how folic acid upregulates DHFR expression
    • Tissue specificity of the H4B-regeneration role not delineated
  10. 2015 High

    Tested whether fast protein motions drive the chemical step, showing hydride transfer in the bacterial ortholog is mass-independent and uncoupled from femtosecond-picosecond motions.

    Evidence Heavy-enzyme (13C/15N/2H) kinetics, kinetic isotope effects, and transition path sampling in ecDHFR

    PMID:26652185

    Open questions at the time
    • Performed in E. coli ortholog; human enzyme not tested
    • Does not exclude slower conformational coupling to product release
  11. 2018 Medium

    Identified a conserved residue (Asp122) tuning catalytically productive flexibility, establishing a Goldilocks principle whereby both excess and deficient coupled motion impair hydride transfer.

    Evidence QM/MM, molecular dynamics, correlated-motion analysis, and kinetics of Asp122 mutants in ecDHFR

    PMID:30040418

    Open questions at the time
    • Primarily in silico; mutant kinetics not validated in vitro in this study
    • Human DHFR equivalent residue not directly tested
  12. 2011 High

    Connected DHFR loss-of-function to human disease, showing a homozygous mutation reduces enzyme activity/protein and causes megaloblastic anemia and cerebral folate deficiency with gene-dosage effect.

    Evidence DHFR enzyme activity and FMTX-binding assays in patient lymphoblastoid cells, homozygosity mapping, and LC-MS/MS folate profiling

    PMID:21310277

    Open questions at the time
    • Single mutation/family; allelic spectrum not defined
    • Mechanism of selective protein destabilization not resolved
  13. 2021 Medium

    Demonstrated a role for DHFR-dependent one-carbon metabolism in cancer stem-like cell biology, showing inhibition reduces brain tumor-initiating cell self-renewal and tumorigenicity.

    Evidence Methotrexate and EphB ligand inhibition, sphere formation, and cerebral organoid glioma model with four BTIC lines

    PMID:33545223

    Open questions at the time
    • Two inhibition approaches but no genetic DHFR knockout validation
    • Downstream one-carbon metabolites mediating self-renewal not identified
  14. 2023 Medium

    Distinguished DHFR degradation from inhibition, showing MTX-based PROTACs degrade DHFR via proteasome/E3 ligase and yield distinct phenotypes from enzymatic inhibition.

    Evidence PROTAC synthesis, Western blot degradation, proteasome and E3-ligase dependence controls in cancer cell lines

    PMID:37875111

    Open questions at the time
    • Why degradation is less lethal than inhibition is unexplained
    • In vivo efficacy and selectivity not established

Open questions

Synthesis pass · forward-looking unresolved questions
  • How DHFR catalytic, transcriptional, and metabolic regulation integrate in human tissues—and whether the bacterial-ortholog flexibility/Goldilocks mechanisms hold for human DHFR—remains unresolved.
  • Hydride-transfer flexibility findings derive from E. coli ortholog, not validated in human DHFR
  • No structural model of human DHFR regulatory complexes at the promoter
  • Mechanism linking folic acid sensing to DHFR transcriptional/protein upregulation undefined

Mechanism profile

Synthesis pass · controlled-vocabulary classification · explore literature graph →
Molecular activity
GO:0016491 oxidoreductase activity 3 GO:0016740 transferase activity 2
Pathway
R-HSA-1430728 Metabolism 3 R-HSA-74160 Gene expression (Transcription) 2

Evidence

Reading pass · 15 per-paper findings extracted from the source corpus
Year Finding Method Journal Conf PMIDs
1985 Pyrimidine dimer removal (nucleotide excision repair) occurs preferentially in the transcriptionally active DHFR gene compared to the genome overall: >67% of dimers removed from a 14.1 kb DHFR restriction fragment by 26 h post-UV irradiation vs. only ~15% genome-wide, establishing that repair efficiency is coupled to transcriptional activity of the locus. UV-endonuclease-based quantitation of pyrimidine dimers in defined restriction fragments of CHO cells amplified for DHFR Cell High 3838150
2001 Repression of the dhfr promoter during cell cycle withdrawal involves cooperation of two distinct complexes: (1) Sp1-pRb-HDAC1 (TSA-sensitive, histone-deacetylase-dependent) and (2) p130-E2F4-DP-1 at the E2F sites (TSA-insensitive). Co-immunoprecipitation showed HDAC1 and hypophosphorylated pRb associate with Sp1 in serum-starved cells; p130 does not co-precipitate with HDAC1. Upon phosphorylation of pRb/p130 after serum stimulation, Sp1-pRb and p130-E2F interactions are lost while Sp1-HDAC1 persists into S phase. Co-immunoprecipitation, TSA treatment, transfection reporter assays, GAL4 recruitment experiments in CHOC400 cells Molecular and cellular biology High 11158299
1996 Genomic footprinting of the dhfr-rep3 promoter region across the cell cycle showed that Sp1 is constitutively bound to four upstream sites and does not vary during G1-to-S transition; two distinct E2F species bind the overlapping E2F sites with different sequence preferences (TTTGGCGC vs. TTTCGCGC), the latter increasing during G1-to-S transition, indicating it is the cell-cycle-regulated activator of dhfr transcription. High-resolution genomic DNase I footprinting, gel mobility shift assays with sequence-specific probes, cell cycle fractionation Molecular and cellular biology High 8552092
1991 During myogenic terminal differentiation, the decrease in DHFR protein synthesis rate is fully accounted for by the decrease in DHFR mRNA levels (both drop to ~5-6% of proliferative levels with matched kinetics); DHFR mRNA remains polysomal with constant ribosome loading, indicating that translational efficiency is unchanged and post-transcriptional regulation does not contribute to DHFR downregulation in quiescent cells. Metabolic labeling of DHFR synthesis rate, Northern blot, polysome fractionation in differentiated H-αR300T myoblasts with 540 copies of amplified DHFR gene Molecular and cellular biology High 2046674
2009 DHFR mediates folic acid (FA)-induced improvement of endothelial NO and tetrahydrobiopterin (H4B) bioavailability: FA upregulates DHFR expression and activity; DHFR converts dihydrofolic acid to tetrahydrofolate (THF) as demonstrated by HPLC fluorescent assay; siRNA knockdown of DHFR or methotrexate pre-treatment abolishes FA-induced restoration of NO levels in angiotensin II-treated cells, establishing DHFR as a required intermediate in FA-dependent vascular protection. HPLC-fluorescence DHFR activity assay, DHFR siRNA knockdown, methotrexate inhibition, superoxide and NO measurements in endothelial cells and Ang II-infused mice Journal of molecular and cellular cardiology High 19660467
2011 Homozygous DHFR mutation p.Asp153Val causes severely reduced DHFR enzymatic activity and reduced DHFR protein expression (not mRNA), leading to megaloblastic anemia and cerebral folate deficiency with neurologic disease; heterozygous cells show intermediate DHFR activity, confirming a gene-dosage effect. DHFR is required to maintain sufficient CSF and RBC folate levels even when plasma folate is normal. DHFR enzyme activity assay and fluorescein-labeled methotrexate (FMTX) binding in EBV-immortalized lymphoblastoid cells, RT-PCR, protein expression analysis, genome-wide homozygosity mapping, liquid chromatography-tandem MS of folate profile American journal of human genetics High 21310277
2020 In Marfan syndrome mice, TGFβ signaling downregulates DHFR protein, uncoupling eNOS and promoting aortic aneurysm formation via a TGFβ/NOX4/DHFR/eNOS-uncoupling feed-forward loop. Anti-TGFβ treatment restored DHFR abundance and recoupled eNOS; folic acid upregulated endothelial DHFR expression and activity to restore H4B, recouple eNOS, and attenuate aneurysm formation. A NO scavenger (PTIO) reversed FA effects on the TGFβ/NOX4 axis, placing DHFR-mediated NO production upstream of TGFβ suppression. In vivo Fbn1C1039G/+ mouse model, anti-TGFβ antibody treatment, folic acid diet, RNAi in human aortic endothelial cells, DHFR activity assay, H4B/ROS measurements, aortic root measurements Redox biology High 33126053
1999 RNA synthesis from the DHFR gene recovers significantly faster after UV irradiation than can be accounted for by removal of photolesions from the transcribed strand, demonstrating that RNA polymerase II can bypass UV-induced lesions in the DHFR gene prior to their repair. In vivo RNA labeling and nuclear run-on transcription assays at three positions within the DHFR gene in UV-irradiated CHO cells, compared to published photolesion removal kinetics Carcinogenesis Medium 10190552
2015 Hydride transfer by E. coli DHFR (ecDHFR; ortholog informative for mechanism) is independent of protein mass: heavy ecDHFR (uniformly labeled with 13C/15N/2H) does not show altered hydride transfer rate constants in refined kinetic and computational (transition path sampling) experiments, but does show faster substrate dissociation. Fast femtosecond-to-picosecond protein motions are not coupled to the chemical (hydride transfer) step. Transition path sampling simulations, kinetic isotope effect measurements, heavy enzyme studies (13C/15N/2H labeling), pre-steady-state kinetics Biochemistry High 26652185
2018 Asp122 in E. coli DHFR (conserved across the DHFR family) is required for optimal hydride transfer: Asp122X mutations (Asn, Ser, Ala) reduce the hydride transfer rate by perturbing coupled protein motions along the reaction coordinate. D122N and D122S inhibit coupled motion while D122A enhances it, yet all three mutations similarly lower catalytic rate, demonstrating a Goldilocks principle of enzyme flexibility for DHFR catalysis. Computational mutagenesis (QM/MM), molecular dynamics simulations, correlated/principal component motion analysis, enzyme kinetics The journal of physical chemistry. B Medium 30040418
2007 Ligand binding to DHFR produces 'network-bridging effects': systematic residue interaction network analysis and molecular dynamics of native and 19 circularly permuted DHFRs show that ligand binding at the active site causes most interaction network paths to pass through the cofactor, shortening average shortest path lengths. The active site coincides with residues of highest network centrality, and chain cleavage in folding element regions near the active site deactivates DHFR by large perturbations in network properties. Residue interaction network analysis, molecular dynamics simulations of native DHFR and 19 circularly permuted variants, network centrality calculations PLoS computational biology Medium 17571919
2023 MTX-based PROTACs achieve proteasome- and E3 ligase-dependent selective degradation of DHFR in multiple cancer cell lines. Unlike MTX treatment (which increases cellular DHFR protein expression), DHFR-targeting PROTACs produce distinct, less-lethal phenotypes, demonstrating that DHFR degradation has different functional consequences from DHFR inhibition. PROTAC synthesis, cell viability assays, Western blot for DHFR degradation, proteasome inhibitor controls, E3 ligase-dependent activity confirmation in cancer cell lines Cell chemical biology Medium 37875111
2021 DHFR inhibition (by methotrexate or EphB activation with synthetic ligands) reduces the self-renewal capacity and tumorigenic potential of human brain tumor initiating cells (BTIC) both in vitro (sphere formation) and in a cerebral organoid glioma model, establishing DHFR activity as required for BTIC self-renewal in one-carbon metabolism. MTX treatment, EphB synthetic ligand activation, sphere formation assay, cerebral organoid glioma (GLICO) model, four human BTIC lines Cancer letters Medium 33545223
1993 Removal of MMS-induced N-methylpurines (base excision repair) is not preferential in the transcriptionally active DHFR gene compared to a non-transcribed flanking region or mitochondrial DNA; repair rates are similar across all three regions, demonstrating that transcription-coupled repair does not apply to N-methylpurine adducts in the DHFR locus. Alkaline hydrolysis generating strand breaks at AP sites after neutral depurination, quantitation in DHFR gene domain, 3'-flanking region, and mitochondrial DNA in CHO-B11 cells Carcinogenesis Medium 8222061
2000 RIP60, a 15-zinc-finger protein that binds ATT-rich sites in the dhfr replication origin oribeta, forms homodimers that loop a 720 bp DNA region in vitro. Structural dissection showed that zinc finger hands Z1 and Z2 independently bind oribeta sites with different affinities; hand Z2 plus part of a proline-rich region is sufficient for protein multimerization and DNA looping. RIP60 has weak replication enhancer activity in plasmid replication assays. One-hybrid screen for human RIP60 cDNA, gel mobility shift assays, DNase I footprinting, ligation enhancement assay, atomic force microscopy, polyomavirus origin-dependent replication assay Nucleic acids research Medium 10606657

Source papers

Stage 0 corpus · 100 papers · ranked by NIH iCite citations
Year Title Journal Citations PMID
1985 DNA repair in an active gene: removal of pyrimidine dimers from the DHFR gene of CHO cells is much more efficient than in the genome overall. Cell 1141 3838150
2019 DHFR Inhibitors: Reading the Past for Discovering Novel Anticancer Agents. Molecules (Basel, Switzerland) 157 30909399
1989 Dual bidirectional promoters at the mouse dhfr locus: cloning and characterization of two mRNA classes of the divergently transcribed Rep-1 gene. Molecular and cellular biology 120 2674679
2005 Targeting DHFR in parasitic protozoa. Drug discovery today 105 15718161
2007 Common origin and fixation of Plasmodium falciparum dhfr and dhps mutations associated with sulfadoxine-pyrimethamine resistance in a low-transmission area in South America. Antimicrobial agents and chemotherapy 102 17283199
1985 Analysis of the mouse dhfr promoter region: existence of a divergently transcribed gene. Molecular and cellular biology 99 3018531
2009 Mechanistic insights into folic acid-dependent vascular protection: dihydrofolate reductase (DHFR)-mediated reduction in oxidant stress in endothelial cells and angiotensin II-infused mice: a novel HPLC-based fluorescent assay for DHFR activity. Journal of molecular and cellular cardiology 92 19660467
2011 Dihydrofolate reductase deficiency due to a homozygous DHFR mutation causes megaloblastic anemia and cerebral folate deficiency leading to severe neurologic disease. American journal of human genetics 81 21310277
2008 Emergence of a dhfr mutation conferring high-level drug resistance in Plasmodium falciparum populations from southwest Uganda. The Journal of infectious diseases 79 18471065
1996 c-Myc overexpression associated DHFR gene amplification in hamster, rat, mouse and human cell lines. Oncogene 78 8570205
1996 Import of a DHFR hybrid protein into glycosomes in vivo is not inhibited by the folate-analogue aminopterin. The Journal of cell biology 77 8636210
1990 X-ray induction of methotrexate resistance due to dhfr gene amplification. Somatic cell and molecular genetics 70 2122527
2001 Cooperation of E2F-p130 and Sp1-pRb complexes in repression of the Chinese hamster dhfr gene. Molecular and cellular biology 58 11158299
2008 Clinical impact of the methotrexate resistance-associated genes C-MYC and dihydrofolate reductase (DHFR) in high-grade osteosarcoma. Annals of oncology : official journal of the European Society for Medical Oncology 55 18385200
2003 DHFR and DHPS genotypes of Plasmodium falciparum isolates from Gabon correlate with in vitro activity of pyrimethamine and cycloguanil, but not with sulfadoxine-pyrimethamine treatment efficacy. The Journal of antimicrobial chemotherapy 54 12805261
1996 Protein-DNA interactions at the major and minor promoters of the divergently transcribed dhfr and rep3 genes during the Chinese hamster ovary cell cycle. Molecular and cellular biology 52 8552092
2012 New mechanisms for an old drug; DHFR- and non-DHFR-mediated effects of methotrexate in cancer cells. Klinicka onkologie : casopis Ceske a Slovenske onkologicke spolecnosti 50 23581023
2020 Design, Synthesis, and Antimicrobial Evaluation of a New Series of N-Sulfonamide 2-Pyridones as Dual Inhibitors of DHPS and DHFR Enzymes. ACS omega 45 32426597
2015 miR-192 suppresses leptomeningeal dissemination of medulloblastoma by modulating cell proliferation and anchoring through the regulation of DHFR, integrins, and CD47. Oncotarget 44 26506238
2011 Association of MTHFR, MTR, MTRR, RFC1, and DHFR gene polymorphisms with susceptibility to sporadic colon cancer. DNA and cell biology 44 21438757
2007 Ligand binding and circular permutation modify residue interaction network in DHFR. PLoS computational biology 44 17571919
1994 Methotrexate does not block import of a DHFR fusion protein into chloroplasts. Plant molecular biology 43 8111032
2008 Plasmodium vivax dhfr and dhps mutations in isolates from Madagascar and therapeutic response to sulphadoxine-pyrimethamine. Malaria journal 42 18302746
2022 New thiazole-based derivatives as EGFR/HER2 and DHFR inhibitors: Synthesis, molecular modeling simulations and anticancer activity. European journal of medicinal chemistry 41 35964425
2020 Targeting feed-forward signaling of TGFβ/NOX4/DHFR/eNOS uncoupling/TGFβ axis with anti-TGFβ and folic acid attenuates formation of aortic aneurysms: Novel mechanisms and therapeutics. Redox biology 41 33126053
2022 In Vitro Antimicrobial Evaluation, Single-Point Resistance Study, and Radiosterilization of Novel Pyrazole Incorporating Thiazol-4-one/Thiophene Derivatives as Dual DNA Gyrase and DHFR Inhibitors against MDR Pathogens. ACS omega 38 35187315
2019 Antimalarial Drug Pyrimethamine Plays a Dual Role in Antitumor Proliferation and Metastasis through Targeting DHFR and TP. Molecular cancer therapeutics 38 30642883
2016 Much more than you expected: The non-DHFR-mediated effects of methotrexate. Biochimica et biophysica acta. General subjects 38 27993660
2007 Variation and expression of dihydrofolate reductase (DHFR) in relation to spina bifida. Molecular genetics and metabolism 38 17336564
1990 Cell cycle-dependent DHFR and t-PA production in cotransfected, MTX-amplified CHO cells revealed by dual-laser flow cytometry. Experimental cell research 38 2110526
2021 Sulfaguanidine Hybrid with Some New Pyridine-2-One Derivatives: Design, Synthesis, and Antimicrobial Activity against Multidrug-Resistant Bacteria as Dual DNA Gyrase and DHFR Inhibitors. Antibiotics (Basel, Switzerland) 37 33562582
2011 Stable expression of green fluorescent protein and targeted disruption of thioredoxin peroxidase-1 gene in Babesia bovis with the WR99210/dhfr selection system. Molecular and biochemical parasitology 37 22108434
2008 dhfr and dhps genotype and sulfadoxine-pyrimethamine treatment failure in children with falciparum malaria in the Democratic Republic of Congo. Tropical medicine & international health : TM & IH 37 19055622
1992 Effect of amplification of dhfr and lac Z genes on growth and beta-galactosidase expression in suspension cultures of recombinant CHO cells. Cytotechnology 36 1369176
1994 Increased methotrexate resistance and dhfr gene amplification as a consequence of induced Ha-ras expression in NIH 3T3 cells. Cancer research 33 8162600
1990 Chromosomal rearrangements and point mutations in the DHFR-TS gene of Plasmodium chabaudi under antifolate selection. Molecular and biochemical parasitology 32 2233898
2000 Expression of carbamoyl phosphate synthetase I and ornithine transcarbamoylase genes in Chinese hamster ovary dhfr-cells decreases accumulation of ammonium ion in culture media. Journal of biotechnology 31 10989172
2015 Hydride Transfer in DHFR by Transition Path Sampling, Kinetic Isotope Effects, and Heavy Enzyme Studies. Biochemistry 30 26652185
1993 Methyl methanesulfonate adduct formation and repair in the DHFR gene and in mitochondrial DNA in hamster cells. Carcinogenesis 30 8222061
2022 New antileishmanial quinoline linked isatin derivatives targeting DHFR-TS and PTR1: Design, synthesis, and molecular modeling studies. European journal of medicinal chemistry 28 36493614
2020 Discovery of New Schiff Bases Tethered Pyrazole Moiety: Design, Synthesis, Biological Evaluation, and Molecular Docking Study as Dual Targeting DHFR/DNA Gyrase Inhibitors with Immunomodulatory Activity. Molecules (Basel, Switzerland) 28 32498469
2017 Synthesis, biological evaluation and molecular modeling study of new (1,2,4-triazole or 1,3,4-thiadiazole)-methylthio-derivatives of quinazolin-4(3H)-one as DHFR inhibitors. Bioorganic chemistry 27 28499189
2019 DHFR/TYMS are positive regulators of glioma cell growth and modulate chemo-sensitivity to temozolomide. European journal of pharmacology 26 31542479
2017 DHFR and MDR1 upregulation is associated with chemoresistance in osteosarcoma stem-like cells. Oncology letters 26 28693150
2008 An insertion/deletion polymorphism of the dihydrofolate reductase (DHFR) gene is associated with serum and red blood cell folate concentrations in women. Human genetics 25 18247058
2023 DihydrofolateReductase (DHFR) Inhibitors: A Comprehensive Review. Current medicinal chemistry 23 36896902
2023 Design, synthesis, anticancer, and antibacterial evaluation of some quinazolinone-based derivatives as DHFR inhibitors. Drug development research 23 37052308
2016 Identifying antimalarial compounds targeting dihydrofolate reductase-thymidylate synthase (DHFR-TS) by chemogenomic profiling. International journal for parasitology 23 27150044
2012 Fusion of the Dhfr/Mtx and IR/MAR gene amplification methods produces a rapid and efficient method for stable recombinant protein production. PloS one 23 23300841
2000 The dhfr oribeta-binding protein RIP60 contains 15 zinc fingers: DNA binding and looping by the central three fingers and an associated proline-rich region. Nucleic acids research 23 10606657
2021 Inhibition of DHFR targets the self-renewing potential of brain tumor initiating cells. Cancer letters 22 33545223
2018 Structural comparison of Mtb-DHFR and h-DHFR for design, synthesis and evaluation of selective non-pteridine analogues as antitubercular agents. Bioorganic chemistry 22 29986181
2016 Trypanosoma brucei DHFR-TS Revisited: Characterisation of a Bifunctional and Highly Unstable Recombinant Dihydrofolate Reductase-Thymidylate Synthase. PLoS neglected tropical diseases 22 27175479
2023 Design, synthesis and antitumor evaluation of novel pyrazolo[3,4-d]pyrimidines incorporating different amino acid conjugates as potential DHFR inhibitors. Journal of enzyme inhibition and medicinal chemistry 21 36382444
2008 Modelling and informatics in the analysis of P. falciparum DHFR enzyme inhibitors. Current medicinal chemistry 21 18673224
1999 Recovery of RNA synthesis from the DHFR gene following UV-irradiation precedes the removal of photolesions from the transcribed strand. Carcinogenesis 20 10190552
2017 Low prevalence of DHFR and DHPS mutations in Pneumocystis jirovecii strains obtained from a German cohort. Infection 19 28303545
2010 Probing the structure of Leishmania donovani chagasi DHFR-TS: comparative protein modeling and protein-ligand interaction studies. Journal of molecular modeling 19 20174846
2010 Improved recombinant protein yield using a codon deoptimized DHFR selectable marker in a CHEF1 expression plasmid. Biotechnology progress 19 20949444
2019 New imidazolone derivatives comprising a benzoate or sulfonamide moiety as anti-inflammatory and antibacterial inhibitors: Design, synthesis, selective COX-2, DHFR and molecular-modeling study. Bioorganic chemistry 18 31796216
2016 Non-DHFR-mediated effects of methotrexate in osteosarcoma cell lines: epigenetic alterations and enhanced cell differentiation. Cancer cell international 18 26929741
2021 3D-QSAR, molecular docking, DFT and ADMET studies on quinazoline derivatives to explore novel DHFR inhibitors. Journal of biomolecular structure & dynamics 17 34825630
2023 Methotrexate-based PROTACs as DHFR-specific chemical probes. Cell chemical biology 16 37875111
2021 Recent Design and Structure-Activity Relationship Studies on the Modifications of DHFR Inhibitors as Anticancer Agents. Current medicinal chemistry 16 31622199
2019 Non-antibiotic Small-Molecule Regulation of DHFR-Based Destabilizing Domains In Vivo. Molecular therapy. Methods & clinical development 16 31649953
2013 Alterations in the expression pattern of MTHFR, DHFR, TYMS, and SLC19A1 genes after treatment of laryngeal cancer cells with high and low doses of methotrexate. Tumour biology : the journal of the International Society for Oncodevelopmental Biology and Medicine 15 23838799
2013 Exploring novel strategies for AIDS protozoal pathogens: α-helix mimetics targeting a key allosteric protein-protein interaction in C. hominis TS-DHFR. MedChemComm 15 24324854
1993 Selective expression of methotrexate-resistant dihydrofolate reductase (DHFR) activity in mice transduced with DHFR retrovirus and administered methotrexate. The Journal of pharmacology and experimental therapeutics 15 8246174
2022 Design, Microwave-Assisted Synthesis, Antimicrobial and Anticancer Evaluation, and In Silico Studies of Some 2-Naphthamide Derivatives as DHFR and VEGFR-2 Inhibitors. ACS omega 14 36157776
2017 Understanding the molecular mechanism of substrate channeling and domain communication in protozoal bifunctional TS-DHFR. Protein engineering, design & selection : PEDS 14 28338744
2015 Frequencies of dhfr/dhps multiple mutations and Plasmodium falciparum submicroscopic gametocyte carriage in Gabonese pregnant women following IPTp-SP implementation. Acta parasitologica 14 26203988
2012 Molecular diagnosis and detection of Pneumocystis jirovecii DHPS and DHFR genotypes in respiratory specimens from Colombian patients. Diagnostic microbiology and infectious disease 14 22321995
2011 Production and characterization of chimeric monoclonal antibodies against Burkholderia pseudomallei and B. mallei using the DHFR expression system. PloS one 14 21573027
2009 Identification and analysis of specific chromosomal region adjacent to exogenous Dhfr-amplified region in Chinese hamster ovary cell genome. Journal of bioscience and bioengineering 14 20347775
2023 New Thiophenyl-pyrazolyl-thiazole Hybrids as DHFR Inhibitors: Design, Synthesis, Antimicrobial Evaluation, Molecular Modeling, and Biodistribution Studies. ACS omega 13 37901585
2019 Maternal Haplotypes in DHFR Promoter and MTHFR Gene in Tuning Childhood Acute Lymphoblastic Leukemia Onset-Latency: Genetic/Epigenetic Mother/Child Dyad Study (GEMCDS). Genes 13 31443485
2016 Low prevalence of dihydro folate reductase (dhfr) and dihydropteroate synthase (dhps) quadruple and quintuple mutant alleles associated with SP resistance in Plasmodium vivax isolates of West Bengal, India. Malaria journal 13 27485211
2015 High prevalence of dhfr and dhps molecular markers in Plasmodium falciparum in pregnant women of Nchelenge district, Northern Zambia. Malaria journal 13 25943379
2013 Evidence of the involvement of the DHFR gene in nonsyndromic cleft lip with or without cleft palate. European journal of medical genetics 13 24361572
2010 High-throughput genotyping of single nucleotide polymorphisms in the Plasmodium falciparum dhfr gene by asymmetric PCR and melt-curve analysis. Journal of clinical microbiology 13 20631115
1991 Changes in dihydrofolate reductase (DHFR) mRNA levels can account fully for changes in DHFR synthesis rates during terminal differentiation in a highly amplified myogenic cell line. Molecular and cellular biology 13 2046674
2018 Effect of Asp122 Mutation on the Hydride Transfer in E. coli DHFR Demonstrates the Goldilocks of Enzyme Flexibility. The journal of physical chemistry. B 12 30040418
2017 DNA variants in DHFR gene and response to treatment in children with childhood B ALL: revisited in AIEOP-BFM protocol. Pharmacogenomics 12 29210328
2023 New 2-alkoxycyanopyridine derivatives as inhibitors of EGFR, HER2, and DHFR: Synthesis, anticancer evaluation, and molecular modeling studies. Bioorganic chemistry 11 37769524
2018 Association of FOLH1, DHFR, and MTHFR gene polymorphisms with susceptibility of Neural Tube Defects: A case control study from Eastern India. Birth defects research 11 30120883
2010 TS, DHFR and GARFT expression in non-squamous cell carcinoma of NSCLC and malignant pleural mesothelioma patients treated with pemetrexed. Anticancer research 11 21036757
2006 DHFR and MSH3 co-amplification in childhood acute lymphoblastic leukaemia, in vitro and in vivo. Carcinogenesis 11 17148505
2024 In Silico and In vitro Analysis of Phenolic Acids for Identification of Potential DHFR Inhibitors as Antimicrobial and Anticancer Agents. Current protein & peptide science 10 37641992
2024 Novel tetrahydroisoquinolines as DHFR and CDK2 inhibitors: synthesis, characterization, anticancer activity and antioxidant properties. BMC chemistry 10 38365746
2024 Dynamic genomic changes in methotrexate-resistant human cancer cell lines beyond DHFR amplification suggest potential new targets for preventing drug resistance. British journal of cancer 10 38594370
2022 New quinazolinone-based derivatives as DHFR/EGFR-TK inhibitors: Synthesis, molecular modeling simulations, and anticancer activity. Archiv der Pharmazie 10 36257809
2017 Extended Linkers Improve the Detection of Protein-protein Interactions (PPIs) by Dihydrofolate Reductase Protein-fragment Complementation Assay (DHFR PCA) in Living Cells. Molecular & cellular proteomics : MCP 10 29203496
2016 Repositioning of DHFR Inhibitors. Current topics in medicinal chemistry 10 26881719
1990 Transfection of DHFR- and DHFR+ mammalian cells using methotrexate-resistant mutants of mouse dihydrofolate reductase. FEBS letters 10 2401371
2023 DHFR Inhibitors Display a Pleiotropic Anti-Viral Activity against SARS-CoV-2: Insights into the Mechanisms of Action. Viruses 9 37243214
2022 Disparate selection of mutations in the dihydrofolate reductase gene (dhfr) of Plasmodium ovale curtisi and P. o. wallikeri in Africa. PLoS neglected tropical diseases 9 36469541
2019 Establishment of DHFR-deficient HEK293 cells for high yield of therapeutic glycoproteins. Journal of bioscience and bioengineering 9 31031194
2015 The 19-bp deletion polymorphism of dihydrofolate reductase (DHFR) and nonsyndromic cleft lip with or without cleft palate: evidence for a protective role. Journal of applied oral science : revista FOB 9 26221921
2005 Concurrence of Plasmodium falciparum dhfr and crt mutations in northern Ghana. Malaria journal 9 16164748
1994 The isolation and characterization of the gene (dfr1) encoding dihydrofolate reductase (DHFR) in Schizosaccharomyces pombe. Gene 9 8088538

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