MOLECULAR-BASIS OF 3-KETOTHIOLASE DEFICIENCY - IDENTIFICATION OF AN AG TO AC SUBSTITUTION AT THE SPLICE ACCEPTOR SITE OF INTRON-10 CAUSING EXON-11 SKIPPING

3-Ketothiolase deficiency (3KTD) is the result of a deficiency in mitochondrial acetoacetyl-CoA thiolase (T2). The molecular basis of 3KTD was analyzed in a patient (GK10) and his family at the protein, cDNA and gene levels. Protein analyses showed that GK10's T2 protein was undetectable in fibroblasts even with the pulse-protein labeling method and that his parents were carriers of 3KTD. Complementary DNA analyses with PCR showed that T2 cDNA in the patient lacked the normal exon 11 sequence and that his parents were obligatory carriers of the DNA sequence which canceled exon 11. When the PCR-amplified genomic fragments around exon 11 were sequenced, an AG underbar to AC underbar mutation at the 3' splice site of intron 10 was detected. This mutation is presumed to be responsible for exon 11 skipping.