Cytosolic arylamine N-acetyltransferase (NAT) deficiency in the dog and other canids due to an absence of NAT genes.
The purpose of this study was to determine the molecular basis in the dog for an unusual and absolute deficiency in the activity of cytosolic N-acetyltransferase (NAT), an enzyme important for the metabolism of arylamine and hydrazine compounds. NAT activity towards two NAT substrates, p-aminobenzoic acid and sulfamethazine, was undetectable in dog liver cytosol, despite substrate concentrations ranging from 10 microM to 4 mM and a wide range of incubation times. Similarly, no protein immunoreactive to NAT antibody was evident on western blot analysis of canine liver cytosol. Southern blot analysis of genomic DNA from a total of twenty-five purebred and mixed bred dogs, and eight wild canids, probed with a full-length human NAT2 cDNA, suggested an absence of NAT sequences in all canids. Polymerase chain reaction amplification of genomic DNA using degenerate primers designed to mammalian NAT1 and NAT2 consensus sequences generated products of the expected size in human, mouse, rabbit, and cat DNA, but no NAT products in any dog or wild canids. These results support the conclusion that cytosolic NAT deficiency in the domestic dog is due to a complete absence of NAT genes, and that this defect is shared by other canids.
Trepanier
L A
LA
Department of Pharmacology, Cornell University, College of Veterinary Medicine, Ithaca, NY 14853, U.S.A. lat1@cornell.edu
Ray
K
K
Winand
N J
NJ
Spielberg
S P
SP
Cribb
A E
AE
D010129:4-Aminobenzoic Acid / Q000378:metabolism; D000818:Animals; D000835:Animals, Wild; D001191:Arylamine N-Acetyltransferase / Q000172:deficiency* / Q000235:genetics*; D015139:Blotting, Southern; D015153:Blotting, Western; D002415:Cats; D003600:Cytosol / Q000201:enzymology; D004247:DNA / Q000302:isolation & purification; D004285:Dogs / Q000235:genetics* / Q000378:metabolism; D019143:Evolution, Molecular; D006801:Humans; D008099:Liver / Q000201:enzymology*; D051379:Mice; D016133:Polymerase Chain Reaction; D011817:Rabbits; D013418:Sulfamethazine / Q000378:metabolism
D013418:Sulfamethazine; D004247:DNA; D001191:Arylamine N-Acetyltransferase; D010129:4-Aminobenzoic Acid