The hinge deletion allelic variant of porcine IgA results from a mutation at the splice acceptor site in the first C alpha intron.
Recently published genomic and cDNA sequences for porcine IgA suggested that the splice acceptor site in the C alpha 1-C alpha 2 intron was an AA rather than an AG dinucleotide. This possibility was tested in an in vitro HeLa cell splicing system using an RNA substrate corresponding to the genomic DNA with the putative AA splice site. Data indicated that splicing occurred at a cryptic AG site 12 nucleotides into the C alpha 2 domain rather than at the AA site. The possibility that swine B cells could use either site was tested by preparing the cDNAs from 13 different samples representing nine animals and amplifying the segment from the first C alpha 1 nucleotide to nucleotide 532 in C alpha 2 (genomic DNA numbering system). Analysis on a 6% polyacrylamide sequencing gel revealed two polynucleotide products in most samples that differed by the expected 12 nucleotides, suggesting that swine could use both splice sites. Sequence analysis confirmed that the shorter form was spliced at the downstream site and the larger form at the apparent upstream AA site. However, when the genomic DNA from an animal expressing only the longer polynucleotide was cloned and sequenced, the upstream splice acceptor site was AG not AA. Thus the data suggested that porcine IgA occurred in two allelic forms, designated IgAa and IgAb, which differ by an apparent G to A mutation in the last nucleotide of intron 1 resulting in a short-hinged (two amino acids, IgAb) variant, in which the downstream cryptic splice site is used, as well as a "normal-hinged" (six amino acids, IgAa) variant. Evidence that IgAa and IgAb are allelic was confirmed by genotypic analyses of progeny from matings of IgAa/IgAb heterozygotes. Evidence that both transcripts are functional was confirmed by showing that serum IgA levels were similar in animals homozygous for each variant.
Brown
W R
WR
Department of Microbiology, University of Iowa, Iowa City 52242, USA.
Kacskovics
I
I
Amendt
B A
BA
Blackmore
N B
NB
Rothschild
M
M
Shinde
R
R
Butler
J E
JE
D000483:Alleles; D000818:Animals; D001483:Base Sequence; D005260:Female; D005803:Genes, Immunoglobulin; D007070:Immunoglobulin A / Q000097:blood; D007137:Immunoglobulin alpha-Chains / Q000235:genetics*; D008297:Male; D008969:Molecular Sequence Data; D012150:Polymorphism, Restriction Fragment Length; D012326:RNA Splicing; D017384:Sequence Deletion; D013552:Swine / Q000276:immunology*
D007070:Immunoglobulin A; D007137:Immunoglobulin alpha-Chains