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Analysisof Thiamin-Responsive Megaloblastic Anemia Syndrome Pathogenesis by Targeted Gene Disruption

Research Project

Project/Area Number 13671196
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeSingle-year Grants
Section一般
Research Field Metabolomics
Research InstitutionKyoto Prefectural University of Medicine

Principal Investigator

NOSAKA Kazuto  Kyoto Prefectural University of Medicine, Grad. Sch. Med., Associate Professor, 医学研究科, 助教授 (10228314)

Co-Investigator(Kenkyū-buntansha) YAMANISHI Kiyofumi  Mie University, Sch. Med, Associate Professor, 医学部, 助教授 (10182586)
Project Period (FY) 2001 – 2003
Project Status Completed (Fiscal Year 2003)
Budget Amount *help
¥3,700,000 (Direct Cost: ¥3,700,000)
Fiscal Year 2003: ¥1,300,000 (Direct Cost: ¥1,300,000)
Fiscal Year 2002: ¥1,700,000 (Direct Cost: ¥1,700,000)
Fiscal Year 2001: ¥700,000 (Direct Cost: ¥700,000)
Keywordsthiamin / thiamin-responsive megaloblastic anemia / thiamin transporter / thiamin pyrophosphokinase
Research Abstract

Thiamin-responsive megaloblastic anemia (TRMA ; OMIM 249270) is an autosomal recessive disorder defined by the occurrence of megaloblastic anemia, diabetes mellitus, sensorineural deafness, and responding to thiamin treatment. It has been speculated that TRMA is caused by the aberration of one of the two proteins both required for thiamin pyrophosphate synthesis, thiamin pyrophosphokinase (TPK) and thiamin transporter. We and others identified SLC192A as the disease gene that encodes the high affinity thiamin transporter (THTR1).
In this study, we isolated the complementary and genome DNAs of mouse thiamin transporter (mTHTR1). Using 5'-RACE two transcriptional start sites for SLC192A located at -175 and -183 relative to the translation start codon were identified in mouse liver cells. We planed to generate a mouse model of SLC192A deficiency to understand the pathogenesis of TRMA. On the other hand, we also isolated human TPK cDNA (hTPK1) and analyzed the enzymatic properties. Furthermore, we identified the minimal 5'-promoter region required for the expression of hTPK1 gene in HepG2 cells which was found to be encoded in a sequence between -540〜-490,and included a Sp1 binding site. Mutation in Sp1 site in the region led to a decrease in the promoter activity. Using Electrophoretic mobility shift analysis with the nuclear extracts from HepG2 cells and the synthetic 24bp oligonucleotide corresponding to -514〜-491 sequence of hTPK1 promoter region, specific DNA-protein complexes were identified. These results suggested the importance of Sp1 in regulating the hTPK1 expression.

Report

(4 results)
  • 2003 Annual Research Report   Final Research Report Summary
  • 2002 Annual Research Report
  • 2001 Annual Research Report
  • Research Products

    (5 results)

All Other

All Publications (5 results)

  • [Publications] Onozuka M.: "Steady-State Kinetics and Mutational Studies of Recombinant Human Thiamin Pyrophosphokinase."J.Nutr.Sci.Vitaminol.. 49・3. 156-162 (2003)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Onozuka M., Nosaka K.: "Steady-State Kinetics and Mutational Studies of Recombinant Human Thiamin Pyrophosphokinase."J.Nutr.Sci.Vitaminol.. 49(3). 156-162 (2003)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Onozuka M.: "Steady-State Kinetics and Mutational Studies of Recombinant Human Thiamin Pyrophosphokinase."J.Nutr.Sci.Vitaminol.. 49・3. 156-162 (2003)

    • Related Report
      2003 Annual Research Report
  • [Publications] Nosaka K.: "Isolation and Characterization of a Human Thiamin Pyrophosphokinase cDNA."Biochim.Biophys.Acta. 1517・2. 293-297 (2001)

    • Related Report
      2001 Annual Research Report
  • [Publications] 野坂 和人: "チアミン反応性貧血症候群の原因遺伝子-高親和性チアミン輸送タンパク質"ビタミン. 75・12. 577-579 (2001)

    • Related Report
      2001 Annual Research Report

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Published: 2001-04-01   Modified: 2016-04-21  

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