| Project/Area Number |
04454154
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| Research Category |
Grant-in-Aid for General Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Research Field |
General medical chemistry
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| Research Institution | Tokyo Medical and Dental University |
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Principal Investigator |
IKAWA Yoji Tokyo Medical and Dental University School of Medicine, Dept.of Biochemsitry, Professor, 医学部・生化学, 教授 (40085618)
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| Co-Investigator(Kenkyū-buntansha) |
KITAYAMA Hitoshi The Insititure of Physical and Chemical Research (RIKEN) The Tsukuba Life Scienc, ライフサイエンス筑波研究センター, 研究員 (30231286)
YAMAMURA Yasuko Tokyo Medical and Dental University School of Medicine, Dept.of Biochemsitry, As, 医学部・生化学, 助手 (50146809)
SATOH Singo Tokyo Medical and Dental University School of Medicine, Dept.of Biochemsitry, As, 医学部・生化学, 助手 (10143562)
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| Project Period (FY) |
1992 – 1994
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| Project Status |
Completed (Fiscal Year 1994)
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| Budget Amount *help |
¥6,500,000 (Direct Cost: ¥6,500,000)
Fiscal Year 1994: ¥2,100,000 (Direct Cost: ¥2,100,000)
Fiscal Year 1993: ¥2,000,000 (Direct Cost: ¥2,000,000)
Fiscal Year 1992: ¥2,400,000 (Direct Cost: ¥2,400,000)
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| Keywords | tumour suppressor gene / small G protein / signal transduction / erythropoietin receptor / novel Ras function / Krev-1 expression / Pain-increased Krev-1 expression / IL-2受容体 / 増殖抑制遺伝子 |
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| Research Abstract |
To investigate novel physiological functions of K-ras and Krev-1 coding G-proteins, we introduced those genes into a mouse EPOR-expressing T cell line (CTLL-2/EPOR) and we found that K-ras gene induced EPO-dependent cell growth, and Krev-1 induced factor-independent growth. EPOR cDNA introduction conferred IL-3-dependent BAF-BO3 cells of a pre-B cell origin EPO-dependent by inducing a low affinity EPOR (More than 1nM concentration of EPOR was needed for the EPO-dependent growth), whereas EPOR cDNA-introduced CTLL-2 clones after introduction of K-ras, C/ERas 4 and C/ERas 6, were expressing a high affinity EPOR and responding to even 20pM EPO for the growth. The EPO-EPOR complex from EPO-treated CERas 4 cells, immunoprecipitated by anti-EPOR antibody was analyzed by SDS-PAGE and immunoblotting with anti-phosphotyrosine, visualizing a 160kDa tyrosine-phosphorylated protein (p160) as well as EPOR and 125 kDa protein. The latter protein was shown to be JAK3.
The Krev-1 expression analized by Western blotting using a monoclonal antibody against the Krev-1 protein revealed two times higher expression of the protein in the spinal cord. Both Krev-1 and K-Ras proteins were present in the spinal cord of rats at birth, and became localized in the synaptic plasma membrane in 5 days after birth, and Krev-1 protein increased in the dorsal horn of the spinal cord 1 hour after experimental pain treatment.
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