Molecular pathogenesis of the patients with congenital disorder of thrombosis and hemostasis
| Project/Area Number |
10670933
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Hematology
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| Research Institution | YAMAGATA UNIVERSITY |
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Principal Investigator |
HAYASHI Tomihiro School of Medicine, Yamagata University, Assistant Professor, 医学部, 講師 (90228586)
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| Project Period (FY) |
1998 – 1999
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| Project Status |
Completed (Fiscal Year 1999)
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| Budget Amount *help |
¥1,700,000 (Direct Cost: ¥1,700,000)
Fiscal Year 1999: ¥600,000 (Direct Cost: ¥600,000)
Fiscal Year 1998: ¥1,100,000 (Direct Cost: ¥1,100,000)
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| Keywords | hereditary qualitative platelet disorder / congenital deficiencies of coagulation factors / gene analysis / Bernard-Soulier syndrome / gene expression / mutation / splicing abnormality / 凝固第X因子欠乏症 |
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| Research Abstract |
I. Analysis of Bernard-Soulier syndrome : PheィイD155ィエD1 (TTT) to Ser (TCT) replacement in the leucine rich motif (LRM) of the GPIX polypeptide does cause BSS phenotype. We have certified this by means of in vitro transfection studies with plasmid for mutant GPIX and other plasmids for GPIb/IX complex. Mutant GPIX could not increase the surface expression of GPIb-α nor surface expression of GPIX itself.
II. Analysis of coagulation factor X deficiency : We have identified the molecular defect underlying congenital factor X (FX) deficiency. A novel 3-base-pair deletion was observed within intron D of the FX gene. This mutation was not detected in 53 unrelated Japanese (106 alleles) by an allele-specific restriction analysis, indicating a likely cause of the FX deficiency. The deletion resides within a polypyrimidine tract of the acceptor splicing site where U2 snRNP binds to form spliceosomes. This defect could alter the formation of splicesomes, then result in incorrect splicing and decre
… More
ased FX production.
III. Analysis of coagulation factor XI deficiency : We have identified a novel single-base point mutation, a GT->AT transition of the factor XI (FXI) gene at the donor splicing site of its intron J.A reverse transcription-polymerase chain reaction with ectopic RNA revealed that propositus cDNA showed 20-base truncation of the 3'-end of the FXI exon 10, resulting in the frame-shift and generation of premature stop codon at 56-base downstream of the correct exon 10/11 junction. A phenotypic consequence of this mutation was then investigated through mammalian cell expression system. Recombinant plasmids harboring wild type FXI gene did direct the production of the FXI antigen into the medium, but the plasmids lacking 20 bases of the FXI exon 10 failed to produce the antigen while the mRNA expression thereof was unchanged. The mutation described here is novel, and ectopic RNA from peripheral mononuclear cells was first proved to be useful for analyzing the resultant FXI mRNA sequence. Less
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Report
(3 results)
Research Products
(15 results)
