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Background  N-glycolylneuraminic acid (Neu5Gc) is synthesized from its precursor N-acetylneuraminic acid (Neu5Ac) by cytidine-5′-monophospho-N acetylneuraminic acid hydroxylase (CMAH), which is encoded by the  CMAH  gene. Most mammals have both Neu5Gc and Neu5Ac, but humans and ferrets have only Neu5Ac because of loss-of-function mutations. Dogs and cats are polymorphic for Neu5Gc and Neu5Ac expression like cats, in which the  CMAH  gene is responsible for the AB Blood group system. Although the  CMAH  gene has been characterized in many species, not much is known about it in dogs. In this study, we cloned the dog  CMAH  cDNA, and performed mRNA expression analysis of this gene in several organs. We also identified single nucleotide polymorphisms (SNPs) in the  CMAH  gene.    Results  We cloned the 1737-bp open reading frame of the dog  CMAH  gene. This gene consists of at least 14 coding exons and codes for a polypeptide of 578 amino acids and is located on chromosome 35. The amino acid identities of dog  CMAH  with the corresponding sequences from cat, pig, chimpanzee, mouse, and rat were high (89 to 93%). RT-PCR analysis showed that the dog  CMAH  cDNA was expressed in various tissues. We identified four exonic SNPs (three synonymous and one non-synonymous), 11 intronic SNPs, and an indel in 11 dog breeds by analyzing the nucleotide sequences of the 14 exons, including the coding region of  CMAH.  In the genotype of the non-synonymous SNP, c.554 A > G (p.Lys185Arg), in a total of 285 dogs of seven different breeds, the allele G was widely distributed, and the allele A was the most frequent in the Shiba dogs. The dogs expressing Neu5Ac did not carry the loss-of-function deletion of  CMAH  found in humans and ferrets, and it remains unclear whether the point mutations influence the expression of Neu5Ac.    Conclusions  We characterized the canine  CMAH  gene at the molecular level for the first time. The results obtained in this study provide essential information that will help in understanding the molecular roles of the  CMAH  gene in canine erythrocyte antigens.

Molecular characterization of cytidine monophospho-N-acetylneuraminic acid hydroxylase (CMAH) associated with the erythrocyte antigens in dogs