Cloning of a rat gene encoding the histo-blood group A enzyme. Tissue expression of the gene and of the A and B antigens.
The complete coding sequence of a BDIX rat gene homologous to the human ABO gene was determined. Identification of the exon-intron boundaries, obtained by comparison of the coding sequence with rat genomic sequences from data banks, revealed that the rat gene structure is identical to that of the human ABO gene. It localizes to rat chromosome 3 (q11-q12), a region homologous to human 9q34. Phylogenetic analysis of a set of sequences available for the various members of the same gene family confirmed that the rat sequence belongs to the ABO gene cluster. The cDNA was transfected in CHO cells already stably transfected with an alpha1,2fucosyltransferase in order to express H oligosaccharide acceptors. Analysis of the transfectants by flow cytometry indicated that A but not B epitopes were synthesized. Direct assay of the enzyme activity using 2' fucosyllactose as acceptor confirmed the strong UDP-GalNAc:Fucalpha1,2GalalphaGalNAc transferase (Atransferase) activity of the enzyme product and allowed detection of a small UDP-Gal:Fucalpha1,2GalalphaGal transferase (B transferase) activity. The presence of the mRNA and of the A and B antigens was searched in various BDIX rat tissues. There was a general good concordance between the presence of the mRNA and that of the A antigen. Tissue distributions of the A and B antigens in the homozygous BDIX rat strain were largely different, indicating that these antigens cannot be synthesized by alleles of the same gene in this rat inbred strain.
INSERM U419, Institut de Biologie, Nantes, France.
Anne
A
Cailleau-Thomas
Béatrice
B
Le Moullac-Vaidye
Jézabel
J
Rocher
Danièle
D
Bouhours
Claude
C
Szpirer
Jacques
J
Le Pendu
D000017:ABO Blood-Group System / Q000378:metabolism; D000595:Amino Acid Sequence; D000818:Animals; D016466:CHO Cells; D002874:Chromosome Mapping; D003001:Cloning, Molecular; D006224:Cricetinae; D018076:DNA, Complementary / Q000235:genetics; D019143:Evolution, Molecular; D006801:Humans; D008969:Molecular Sequence Data; D017350:N-Acetylgalactosaminyltransferases / Q000096:biosynthesis / Q000235:genetics* / Q000378:metabolism; D009844:Oligosaccharides / Q000096:biosynthesis; D025942:Oligosaccharides, Branched-Chain; D009928:Organ Specificity; D010802:Phylogeny; D012333:RNA, Messenger / Q000096:biosynthesis; D051381:Rats; D011919:Rats, Inbred Strains; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid; D014162:Transfection; D014312:Trisaccharides / Q000096:biosynthesis / Q000378:metabolism
D000017:ABO Blood-Group System; D018076:DNA, Complementary; D009844:Oligosaccharides; D025942:Oligosaccharides, Branched-Chain; D012333:RNA, Messenger; D014312:Trisaccharides; C101335:blood group A trisaccharide; C406478:blood group B trisaccharide; D017350:N-Acetylgalactosaminyltransferases; C003742:UDPgalactosamine-galactose acetylgalactosaminyltransferase; C031420:2'-fucosyllactose