Sequence and organization of a Macaca nemestrina carbonic anhydrase I gene.

To identify the cause of an inherited erythrocyte specific deficiency of carbonic anhydrase (CA) in Macaca nemestrina, pigtail macaques, genomic libraries were constructed with DNA from an individual with two normally-expressed CAI alleles and were screened with a human CAI probe containing exons 6 and 7. Four CAI clones were isolated and subclones were constructed in plasmid sequencing vectors to determine the nucleotide sequence of exons 2 through 7. The intron sizes were determined by restriction mapping and Southern methods. Immunodiffusion studies revealed the presence of CAI in the colons of macaques with erythrocyte CAI deficiencies. Southerns comparing genomic DNA from macaques with normal and deficient levels of erythrocyte CAI showed no detectable restriction fragment size differences 3$\sp\prime$ to exon 2 with available CAI probes. To amplify and sequence Macaca nemestrina CAI exon 1a, primers were synthesized for polymerase chain reactions based on sequences flanking the human exon which is located over 35 kb upstream from exon 1c. Nucleotide sequences in the 5$\sp\prime$ regulatory regions of CAI genes from other species with normal or deficient levels of erythrocyte CAI will be compared in continuing studies of carbonic anhydrase tissue-specific gene expression. In primates at least seven CA isozymes are expressed, each with distinct tissue distributions, subcellular locations, levels of expression, specific activities, amino acid sequence lengths, gene sizes and chromosomal locations, and proposed functions. Inherited red cell CAI deficiencies occur in at least three species: human, pigtail macaque, and chinchilla, where CAI is expressed in tissues other than erythrocytes and the individuals homozygous for the erythrocyte CAI deficiency gene do not exhibit any physiological abnormalities. The erythrocyte CAI deficiency has reached polymorphic levels in natural populations of macaques where homozygosity for the CAI deficiency allele results in trace amounts of erythrocyte CAI as well as a concomitant 60% reduction in erythrocyte CAII. The recent demonstration of tissue-specific promoters and splicing events in human and mouse CAI genes suggest a possible mechanism for the cause of the macaque erythrocyte CAI deficiency, i.e., a mutation in the erythrocyte-specific regulatory region of the CAI gene.PhDHuman GeneticsUniversity of Michigan, Horace H. Rackham School of Graduate Studieshttp://deepblue.lib.umich.edu/bitstream/2027.42/105358/1/9023608.pdfDescription of 9023608.pdf : Restricted to UM users only