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Literature summary for 1.5.1.33 extracted from

  • Bhattacharya, A.; Leprohon, P.; Ouellette, M.
    Combined gene deletion of dihydrofolate reductase-thymidylate synthase and pteridine reductase in Leishmania infantum (2021), PLoS Negl. Trop. Dis., 15, e0009377 .
    View publication on PubMedView publication on EuropePMC
Search for more literature for 1.5.1.33

Protein Variants

Protein Variants Comment Organism
additional information a Leishmania infantum DHFR-TS null mutant is thymidine auxotroph showing a phenotype that can be rescued by the addition of thymidine or by transfection of the flavin-dependent bacterial thymidylate synthase (TS) gene ThyX. In DHFR-TS null mutants, it is impossible to obtain a chromosomal null mutant of PTR1 except if DHFR-TS or PTR1 are provided episomally. Episomal expression of PTR1 in these cells does not revert thymidine auxotrophy. Leishmania can survive without copies of either DHFR-TS or PTR1 but not without both. Provided that the results observed with the insect stage parasites are also replicated with intracellular parasites, it suggests that antifolate therapy in Leishmania only works if both DHFRTS and PTR1 are targeted simultaneously. Generation of a PTR1 null mutant in the DHFR-TSNEO/HYG cells using puromycin (PURO) and zeocin (ZEO) inactivation cassettes. Transfection of the PURO cassette in DHFR-TSNEO/HYG cloned cells leads to PTR1PURO/+ parasites. Transfection of the ZEO cassette in these parasites leads to the correct integration at the PTR1 locus but with a remaining intact copy of PTR1 in the population of cells analyzed. DHFR-TSNEO/HYG PTR1PURO/+ cells are complemented with episomally-expressed ThyX. These cells are able to grow in the absence of thymidine supplementation. Upon the transfection of the ZEO PTR1-inactivation cassette integration at the right locus is ontained but the hybridization signals are consistent with the population of parasites having the PTR1PURO/ZEO/+ genotype. This PTR1PURO/ZEO/+ genotype is also obtained in an independent PTR1 gene knockout experiment where ThyX is episomally expressed. When the genetic complementation is done with DHFR-TS instead of ThyX, the transfection of the ZEO PTR1-inactivation leads to a PTR1 chromosomal null mutant Leishmania infantum

Inhibitors

Inhibitors Comment Organism Structure
methotrexate MTX Leishmania infantum
additional information a Cosseq genome-wide gain of function screen is performed against methotrexate and against the two thymidylate synthase inhibitors 5-fluorouracil and pemetrexed Leishmania infantum

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
7,8-dihydrobiopterin + NADPH + H+ Leishmania infantum
-
 5,6,7,8-tetrahydrobiopterin + NADP+
-
 ?
7,8-dihydrobiopterin + NADPH + H+ Leishmania infantum MHOM/MA/67/ITMAP-263
-
 5,6,7,8-tetrahydrobiopterin + NADP+
-
 ?

Organism

Organism UniProt Comment Textmining
Leishmania infantum A4I067
-
-
Leishmania infantum MHOM/MA/67/ITMAP-263 A4I067
-
-

Source Tissue

Source Tissue Comment Organism Textmining
promastigote cultured at 25°C in SDM-79 or M199 Leishmania infantum
-

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
7,8-dihydrobiopterin + NADPH + H+
-
 Leishmania infantum 5,6,7,8-tetrahydrobiopterin + NADP+
-
 ?
7,8-dihydrobiopterin + NADPH + H+
-
 Leishmania infantum MHOM/MA/67/ITMAP-263 5,6,7,8-tetrahydrobiopterin + NADP+
-
 ?

Synonyms

Synonyms Comment Organism
pteridine reductase 1
-
 Leishmania infantum
PTR1
-
 Leishmania infantum

Cofactor

Cofactor Comment Organism Structure
NADPH
-
 Leishmania infantum

General Information

General Information Comment Organism
malfunction Leishmania can survive without copies of either DHFR-TS or PTR1 but not without both. Leishmania is pterin auxotroph but a PTR1 knockout cell will grow well if sufficient reduced pterins are available Leishmania infantum
metabolism folates are reduced in Leishmania by the bifunctional dihydrofolate reductase thymidylate synthase (DHFR-TS) and by pteridine reductase (PTR1) Leishmania infantum
physiological function PTR1 is essential in the absence of an intact bifunctional dihydrofolate reductase thymidylate synthase (DHFR-TS) even in the presence of flavin-dependent bacterial TS gene ThyX or thymidine supplementation, indicating the essential role of reduced pterins or folate beyond thymidine synthesis Leishmania infantum