MOLECULAR-BASIS FOR TYPE-1 ANTITHROMBIN DEFICIENCY - IDENTIFICATION OF 2 NOVEL POINT MUTATIONS AND EVIDENCE FOR A DE-NOVO SPLICE-SITE MUTATION

Inherited type 1 antithrombin (AT) deficiency is characterized by a re duction in both immunologically and functionally detectable protein. T he disorder is associated with a high risk of thromboembolic disease. We have investigated the molecular basis of type 1 AT deficiency in th ree unrelated families. We have used the polymerase chain reaction sin gle-strand conformation polymorphism (PCR-SSCP) analysis, followed by direct sequencing of the seven exons and the intron-exon junctions of the AT gene. Two novel point mutations were identified. A T to C singl e-base substitution was found in codon 421 in exon 6 (nucleotide posit ion 13380), leading to an AT 421 isoleucine to threonine substitution. In another kindred, one of three Cs at nucleotide (nt) positions 5448 to 5450 in exon 3A (codon 151 or 152) was deleted, resulting in a fra meshift mutation and predicting premature termination of protein trans lation at codon 251. In a third family, a previously reported G to A s ubstitution, at nt position 9788 in intron 4, 14 bp in front of exon 5 , was found. We have demonstrated the creation of a de novo exon 5 spl ice site by ectopic transcript analysis of lymphocyte mRNA. In all cas es, the affected individuals were heterozygous for the mutation and no variant AT protein was detected. (C) 1994 by The American Society of Hematology.