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Molecular mechanisms for the assembly of the red cell membrane skeleton during erythroid cell development

Research Project

Project/Area Number 14360187
Research Category

Grant-in-Aid for Scientific Research (B)

Allocation TypeSingle-year Grants
Section一般
Research Field Applied veterinary science
Research InstitutionHOKKAIDO UNIVERSITY

Principal Investigator

INABA Mutsumi  Hokkaido University, Graduate School of Veterinary Medicine, Professor, 獣医学研究科, 教授 (00183179)

Co-Investigator(Kenkyū-buntansha) TAKAKUWA Yuichi  Tokyo Women's Medical University, Faculty of Medicine, Professor, 医学部, 教授 (40113740)
SAITO Masayuki  Hokkaido University, Graduate School of Veterinary Medicine, Professor, 大学院・獣医学研究科, 教授 (80036441)
MAEDA Yoshimitsu  Hokkaido University, Graduate School of Veterinary Medicine, Professor, 大学院・獣医学研究科, 教授 (40002084)
INANAMI Osamu  Hokkaido University, Graduate School of Veterinary Medicine, Associate Professor, 大学院・獣医学研究科, 助教授 (10193559)
YAMAMOTO Masayuki  Tsukuba University, TARA Center, Professor, 基礎医学系先端学際領域研究センター, 教授 (50166823)
滝口 満喜  北海道大学, 大学院・獣医学研究科, 講師 (70261336)
Project Period (FY) 2002 – 2003
Project Status Completed (Fiscal Year 2003)
Budget Amount *help
¥14,800,000 (Direct Cost: ¥14,800,000)
Fiscal Year 2003: ¥4,900,000 (Direct Cost: ¥4,900,000)
Fiscal Year 2002: ¥9,900,000 (Direct Cost: ¥9,900,000)
Keywordserythroid development / red blood cell / membrane skeleton / band 3 / ankyrin / membrane trafficking / intracellular vesicles / cell culture / 赤芽球系分化 / 赤血球膜 / 膜骨格タンパク質 / スペクトリン / 牛
Research Abstract

The "unit-assembly hypothesis" for red cell membrane skeleton during erythroid development was investigated.
Stable transfectants of major constituents of red cell transmembrane proteins were developed in K562 and HEK293 cells using retroviral vectors. Band 3 and glycophorinCweretransportedtotheplasmamembranevia the ER/Golgi pathway. Band 3-binding domain of ankyrin (AnkN90) and protein 4.1 showed membranous localization in the cytoplasm with co-localization with markers for the ER and the Golgi apparatus in band 3-transfeted cells despite of the difference in erythroid differentiation. Endogenous membrane skeletal proteins including ankyrin, protein 4.1, and spectrin also exhibited membranous distribution. These results indicate that the assembly of membrane skeletal proteins occurs at an early stage of erythroidal development on the surface of the ER but not on the inner surface of the plasma membrane.
Stable expression of band 3 caused a reduction in a proliferation rate of K562 cells to approximately 70% that of non-transfected cells. Reduced production of hemoglobin suggestive of erythroid differentiation was also observed. However, no significant changes in intracellular localization of skeletal proteins and cell morphology was detected Synthesis and assembly to the ER of skeletal proteins involving spectrin and subsequent synthesis and localization to the plasma membrane of band 3 were observed in erythroid precursor cells from peripheral blood of cattle in two phase liquid culture system, indicating that the assembly of membrane skeletal proteins to the ER occurs in physiolosical erythropoiesis.
The present study demonstrated that the assembly of the red cell membrane skeleton is an event which takes place at early stages of erythroid development on the ER. The exact mechanism by which a small unit of the ekeleton formed on intracellular vesicles are integrated in the plasma membrane remained to be clarified.

Report

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

    (21 results)

All Other

All Publications (21 results)

  • [Publications] Takada, K., ほか3名: "Spontaneous development of multiple glandular and extraglandular lesions in aged IQI/Jic mice : a model for primary Sjogren's syndrome"Rheumatology. (in press). (2004)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Tahara, T., ほか7名: "Heme positively regulates the expression of beta-globin at the locus control region via the transcriptional factor Bach1 in erythroid cells"Journal of Biological Chemistry. 279. 5480-5487 (2004)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Saito, K., ほか6名: "In situ observation of mobility and anchoring of PKCb in plasma membrane"FEBS Lett.. 541. 126-131 (2003)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Hossain, M.A., ほか6名: "Serum from dogs infected with Babesia gibsoni inhibits maturation of reticulocytes and erythrocyte 5'-nucleotidase activity invitro"Journal of Veterinary Medical Science. 65. 1281-1286 (2003)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Tamahara, S., ほか5名: "Nonessential roles of cysteine residues in functional expression and redox regulatory pathways for canine glutamate/aspartate transporter based on mutagenic analysis"Biochemical Journal. 367. 107-111 (2002)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Hanspal, M., ほか5名: "Plasmodium falciparum cysteine protease falcilain-2 cleaves erythrocyte membrane skeletal proteins at late stages of parasite development"Blood. 100. 1048-1054 (2002)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Takada, K., Takiguchi, M., Konno, A., Inaba, M.: "Spontaneous development of multiple glandular and extraglandular lessons us aged IQI/Jic mice a model for primary Sjogren's syndrome"Rheumatology. (in press). (2004)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Tahara T, Sun J, Nakamshi K, Yamamoto M, Mori H, Saito T, Fujita H, Igarashi K, Taketani S: "Heme positively regulates the expression of beta-globin at the locus control region via the transcriptional factor Bach1 us erythroid cells"J.Biol.Chem.. 279. 5480-5487 (2004)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Saito K, Ito E, Takakuwa Y, Tamura M, Kinjo M.: "In situ observation of mobility and anchoring of PKCbetaI in plasma membrane"FEBS Lett.. 541. 126-131 (2003)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Hossain MA, Yamato O, Yamasaki M, Jeong JR, Chang HS, Maede Y.: "Serum from dogs Infected with Babesia gibsoni inhibits maturation of reticulocytes and erythrocyte 5'-nucleotidase activity in vitro"J.Vet.Med.Sci.. 65. 1281-1286 (2003)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Tamahara, S., Inaba, M., Sato, K., Matsuki, N., Hikasa, Y., Ono, K.: "Nonessential roles of cysteine residues in functional expression and redox regulatory pathways for canine glutamate/aspartate transporter based on mutagenic analysis"Biochem.J.. 367. 107-111 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Hanspal M, Dua M, Takakuwa Y, Chishti AH, Mizuno A.: "Plasmodium falciparum cysteine protease falcipain-2 cleaves erythrocyte membrane skeletal proteins at late stages of parasite development"Blood. 100. 1048-1054 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Tamahara, S.ら6名: "Non-essential roles of cysteine residues in functional expression and redox regulatory pathways for canine glutamate/aspartate transporter based on mutagenic analysis."Biochemical Journal. 367. 107-111 (2002)

    • Related Report
      2003 Annual Research Report
  • [Publications] Thongsong, B.ら7名: "Effects of insulin-like growth factor-I on maternal and fetal plasma amino acid levels in pregnant rats."Journal of Veterinary Medical Sciences. 64. 859-861 (2002)

    • Related Report
      2003 Annual Research Report
  • [Publications] Hanspal, M.ら5名: "Plasmodium falciparum cysteine protease falcilain-2 cleaves erythrocyte membrane skeletal proteins at late stages of parasite development."Blood. 100. 1048-1054 (2002)

    • Related Report
      2003 Annual Research Report
  • [Publications] Saito, K.ら6名: "In situ observation of mobility and anchoring of PKCβ in plasma membrane."FEBS Letter. 541. 126-131 (2003)

    • Related Report
      2003 Annual Research Report
  • [Publications] Hossain, M.A.ら6名: "Serum from dogs infected with Babesia gibsoni inhibits maturation of reticulocytes and erythrocyte 5'-nucleotidase activity invitro."Journal of Veterinary Medical Sciences. 65. 1281-1286 (2003)

    • Related Report
      2003 Annual Research Report
  • [Publications] Tahara, T.ら6名: "Heme positively regulates the expression of beta-globin at the locus control region via the transcriptional factor Bach1 in erythroid cells."Journal of Biological Chemistry. 279. 5480-5487 (2004)

    • Related Report
      2003 Annual Research Report
  • [Publications] Tamahara, S., ほか5名: "Nonessential roles of cysteine residues in functional expression and redox regulatory pathways for canine glutamate/aspartate transporter based on mutagenic analysis"Biochemical Journal. 367. 107-111 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] Matsuki, N., ほか2名: "Catabolism of cytoplasmic and intramitochondrial adenosine nucleotides in C_2C_<12> skeletal myotube under chemical hypoxia"The Journal of Veterinary Medical Science. 64. 341-347 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] Manno, S., ほか2名: "Identification of a functional role for lipid asymmetry in biological membranes : Phosphatidylserine-skeletal protein interactions modulate membrane stability"Proceedings of the National Academy of Sciences of the U.S.A.. 99. 1943-1948 (2002)

    • Related Report
      2002 Annual Research Report

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Published: 2002-04-01   Modified: 2016-04-21  

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