Dihydropteridine reductase as an alternative to dihydrofolate reductase for synthesis of tetrahydrofolate in Thermus thermophilus.
A strategy devised to isolate a gene coding for a dihydrofolate reductase from Thermus thermophilus DNA delivered only clones harboring instead a gene (the T. thermophilus dehydrogenase [DH(Tt)] gene) coding for a dihydropteridine reductase which displays considerable dihydrofolate reductase activity (about 20% of the activity detected with 6,7-dimethyl-7,8-dihydropterine in the quinonoid form as a substrate). DH(Tt) appears to account for the synthesis of tetrahydrofolate in this bacterium, since a classical dihydrofolate reductase gene could not be found in the recently determined genome nucleotide sequence (A. Henne, personal communication). The derived amino acid sequence displays most of the highly conserved cofactor and active-site residues present in enzymes of the short-chain dehydrogenase/reductase family. The enzyme has no pteridine-independent oxidoreductase activity, in contrast to Escherichia coli dihydropteridine reductase, and thus appears more similar to mammalian dihydropteridine reductases, which do not contain a flavin prosthetic group. We suggest that bifunctional dihydropteridine reductases may be responsible for the synthesis of tetrahydrofolate in other bacteria, as well as archaea, that have been reported to lack a classical dihydrofolate reductase but for which possible substitutes have not yet been identified.
Wilquet
Valérie
V
Microbiology, Université Libre de Bruxelles, B-1070 Brussels, Belgium.
Van de Casteele
Mark
M
Gigot
Daniel
D
Legrain
Christianne
C
Glansdorff
Nicolas
N
D000595:Amino Acid Sequence; D004093:Dihydropteridine Reductase / Q000502:physiology*; D008969:Molecular Sequence Data; D013194:Staining and Labeling; D013762:Tetrahydrofolate Dehydrogenase / Q000032:analysis / Q000235:genetics / Q000502:physiology*; D013763:Tetrahydrofolates / Q000096:biosynthesis*; D016963:Thermus thermophilus / Q000235:genetics / Q000378:metabolism*
D013763:Tetrahydrofolates; C030371:5,6,7,8-tetrahydrofolic acid; D013762:Tetrahydrofolate Dehydrogenase; D004093:Dihydropteridine Reductase