| Project/Area Number |
12460137
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| Research Category |
Grant-in-Aid for Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Applied veterinary science
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| Research Institution | The University of Tokyo |
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Principal Investigator |
INABA Mutsumi Graduate School of Agriculture and Life Sciences, The University of Tokyo, Asssociate Professor, 大学院・農学生命科学研究科, 助教授 (00183179)
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| Co-Investigator(Kenkyū-buntansha) |
TAKAKUWA Yuichi Tokyo Women's Medical University, Professor, 医学部, 教授 (40113740)
ONO Ken-ichiro Graduate School of Agriculture and Life Sciences, The University of Tokyo, Professor, 大学院・農学生命科学研究科, 教授 (50111480)
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)
MATSUKI Naoaki Graduate School of Agriculture and Life Sciences, The University of Tokyo, Research Associate, 大学院・農学生命科学研究科, 助手 (40251417)
辻本 元 東京大学, 大学院・農学生命科学研究科, 教授 (60163804)
稲波 修 北海道大学, 大学院・獣医学研究科, 助教授 (10193559)
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| Project Period (FY) |
2000 – 2001
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| Project Status |
Completed (Fiscal Year 2001)
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| Budget Amount *help |
¥14,300,000 (Direct Cost: ¥14,300,000)
Fiscal Year 2001: ¥3,600,000 (Direct Cost: ¥3,600,000)
Fiscal Year 2000: ¥10,700,000 (Direct Cost: ¥10,700,000)
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| Keywords | red cell membranes / membrane skeleton / band 3 / ankyrin / congenital hemolytic anemia / proeasome / vesicle trafficking / cattle / 遺伝性溶血性貧 / プロテアソーム / 膜蛋白質 / 膜移行 / 遺伝性バンド3欠損症 |
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| Research Abstract |
Stable transfectants of normal bovine band 3 (bebWT) or the mutant band 3 with R664X mutation (bebRX) were established in K562 cells and HEK293 cells using retoriviral vectors. Transfected cells expressing EGFP-bebWT and EGFP-bebRX, and N-terminal domain of ankyrin (AnkN90) in combination with band 3 proteins were also prepared.
The bebWT and EGFP-bebWT showed stable expression on the plasma membrane of the transfected cells, whereas the mutant proteins, bebRX and EGFP-bebRX were degraded by Ub-proteasome system soon after synthesis on the ER or after retrograde transported from the Golgi apparatus to the ER. Stability of the bebWT was extremely reduced when bebRX was co-transfected. AnkN90 showed membrane localization within the cells and was destabilized in the cells that had the mutant band 3.
These findings indicate that the mutant band 3 (bebRX) plays a dominant-negative role on the expression of normal band 3 and a partner in the membrane skeleton, ankyrin, and the interaction of band 3 with ankyrin occurs on the ER membrane soon after band 3 synthesis is started during erythroid development.
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