cDNA cloning and chromosome mapping of human dihydropyrimidine dehydrogenase, an enzyme associated with 5-fluorouracil toxicity and congenital thymine uraciluria.
The pig and human dihydropyrimidine dehydrogenase (DPD) cDNAs were cloned and sequenced. The pig enzyme, expressed in Escherichia coli, catalyzed the reduction of uracil, thymine, and 5-fluorouracil with kinetics approximating those published for the enzyme purified from mammalian liver. DPD could be expressed in significant quantities only when uracil was added to the bacterial growth medium. The pig and human enzymes contained 1025 amino acids and calculated M(r) = 111,416 and 111,398, respectively. Conserved domains corresponding to a possible NADPH binding site and FAD binding site were found in the NH2-terminal half of the proteins and two motifs of putative [4Fe-4S] binding sites were found near to the carboxyl terminus of the enzyme. The latter corresponds to the labile COOH-terminal fragment previously shown to contain the iron sulfur centers. A sequence encompassing a peptide corresponding to the uracil binding site was found between the NADPH/FAD-containing NH2-terminal portion of the protein and the iron-sulfur binding sites near to the COOH terminus. Thus, the DPD appears to be derived from at least three distinct domains. The DPYD gene was localized to the centromeric region of human chromosome 1 between 1p22 and q21.
Yokota
H
H
Laboratory of Molecular Carcinogenesis, National Cancer Institute, Bethesda, Maryland 20892.
Fernandez-Salguero
P
P
Furuya
H
H
Lin
K
K
McBride
O W
OW
Podschun
B
B
Schnackerz
K D
KD
Gonzalez
F J
FJ
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002874:Chromosome Mapping; D002878:Chromosomes, Human, Pair 1; D006224:Cricetinae; D018076:DNA, Complementary; D042943:Dihydrouracil Dehydrogenase (NADP); D005472:Fluorouracil / Q000009:adverse effects*; D006801:Humans; D006822:Hybrid Cells; D008661:Metabolism, Inborn Errors / Q000235:genetics*; D051379:Mice; D008969:Molecular Sequence Data; D010088:Oxidoreductases / Q000172:deficiency / Q000235:genetics*; D011994:Recombinant Proteins / Q000235:genetics; D013552:Swine; D013941:Thymine / Q000652:urine*; D014498:Uracil / Q000652:urine*
D018076:DNA, Complementary; D011994:Recombinant Proteins; D014498:Uracil; D010088:Oxidoreductases; D042943:Dihydrouracil Dehydrogenase (NADP); D013941:Thymine; D005472:Fluorouracil