title: Identification of a novel amino acid deletion mutation and a very rare single nucleotide variant in a Japanese family with type I antithrombin deficiency.
abstract: We studied a Japanese family with type I antithrombin (AT) deficiency and identified a novel in-frame deletion mutation (-ATG at nucleotide position of 2771-2773) in the AT gene, which predicted loss of a methionine (Met) at amino acid number of 103. In addition, we found a single base replacement of G to A at nucleotide position of 67 (4 base upstream to the initial codon) in the mutant allele. Since the G67A substitution in the AT gene was very rare, this family was the second case, in which the nucleotide change was transmitted. To elucidate the mechanism of AT deficiency, we transiently expressed wild type and the mutant AT (DeltaM103) in HuH-7 human hepatoma cells and performed pulse-chase studies. The experiments revealed that the mutant AT (DeltaM103) hardly secreted into the medium and underwent partial intracellular degradation. In addition, we performed luciferase reporter assay to examine the effect of G67A substitution on the AT gene expression, and found that the substitution did not reduce the luciferase activity. These results suggested that secretion defect and intracellular degradation of the variant molecule with the deletion of Met 103 were responsible for AT deficiency in this family.
authors.affiliation: Department of Internal Medicine, Ise Municipal Hospital, Mie, Japan.
authors.forename: Kan
authors.initials: K
authors.lastname: Katayama
authors.forename: Natsuko
authors.initials: N
authors.lastname: Hashimoto
authors.forename: Yuki
authors.initials: Y
authors.lastname: Tanaka
authors.forename: Tetsuo
authors.initials: T
authors.lastname: Ozawa
authors.forename: Yoshiharu
authors.initials: Y
authors.lastname: Emi
authors.forename: Takeshi
authors.initials: T
authors.lastname: Ikeda
authors.forename: Miyuki
authors.initials: M
authors.lastname: Katayama
authors.forename: Shinsuke
authors.initials: S
authors.lastname: Nomura
authors.forename: Isao
authors.initials: I
authors.lastname: Kitajima
authors.forename: Takeshi
authors.initials: T
authors.lastname: Nakano
authors.forename: Tsuneo
authors.initials: T
authors.lastname: Imanaka
mesh_terms: D000328:Adult; D000990:Antithrombin III / Q000235:genetics / Q000378:metabolism; D020152:Antithrombin III Deficiency / Q000235:genetics*; D001692:Biological Transport; D025861:Blood Coagulation Disorders, Inherited / Q000235:genetics; D045744:Cell Line, Tumor; D003001:Cloning, Molecular; D004252:DNA Mutational Analysis; D005192:Family Health; D005786:Gene Expression Regulation / Q000235:genetics; D006801:Humans; D007564:Japan; D008297:Male; D010375:Pedigree; D020641:Polymorphism, Single Nucleotide; D017384:Sequence Deletion; D014162:Transfection
chemical_list: D000990:Antithrombin III