Cloning and characterization of the human 5,10-methenyltetrahydrofolate synthetase-encoding cDNA.
Methenyltetrahydrofolate synthetase (MTHFS) catalyses the obligatory initial metabolic step in the intracellular conversion of 5-formyltetrahydrofolate to other reduced folates. We have isolated and sequenced a human MTHFS cDNA which is 872-bp long and codes for a 203-amino-acid protein of 23,229 Da. Escherichia coli BL21(DE3), transfected with pET11c plasmids containing an open reading frame encoding MTHFS, showed a 100-fold increase in MTHFS activity in bacterial extracts after IPTG induction. Northern blot studies of human tissues determined that the MTHFS mRNA was expressed preferentially in the liver and Southern blot analysis of human genomic DNA suggested the presence of a single-copy gene.
Centre de Recherche, Hôtel-Dieu de Montréal, Québec, Canada.
A
A
Dayan
R
R
Bertrand
M
M
Beauchemin
D
D
Chahla
A
A
Mamo
M
M
Filion
D
D
Skup
B
B
Massie
J
J
Jolivet
D000595:Amino Acid Sequence; D001483:Base Sequence; D019731:Carbon-Nitrogen Ligases; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000235:genetics*; D004926:Escherichia coli / Q000235:genetics; D018628:Gene Dosage; D006801:Humans; D008025:Ligases / Q000737:chemistry / Q000235:genetics*; D008099:Liver / Q000737:chemistry; D008969:Molecular Sequence Data; D008970:Molecular Weight; D016366:Open Reading Frames / Q000235:genetics; D012333:RNA, Messenger / Q000032:analysis* / Q000096:biosynthesis; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000737:chemistry; D017422:Sequence Analysis, DNA; D017386:Sequence Homology, Amino Acid
D018076:DNA, Complementary; D012333:RNA, Messenger; D011993:Recombinant Fusion Proteins; D008025:Ligases; D019731:Carbon-Nitrogen Ligases; C021557:5,10-methenyltetrahydrofolate synthetase