| Project/Area Number |
10670521
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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 |
Gastroenterology
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| Research Institution | Kurume University |
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Principal Investigator |
SHICHIJO Shigeki Kurume Univ., Sch. Med., Dep. Immunol., Associate Prof., 医学部, 助教授 (30080592)
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| Co-Investigator(Kenkyū-buntansha) |
ITOH Kyogo Kurume Univ., Sch. Med., Dep. Immunol., Prof., 医学部, 教授 (50125499)
IMAI Yasuhisa Kurume Univ., Sch. Med., Department of lmmunol., Assistant, 医学部, 助手 (90268847)
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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 |
¥3,400,000 (Direct Cost: ¥3,400,000)
Fiscal Year 1999: ¥1,100,000 (Direct Cost: ¥1,100,000)
Fiscal Year 1998: ¥2,300,000 (Direct Cost: ¥2,300,000)
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| Keywords | Tumor rejection antigen / DNA-binding protein / Cell cycle / Apoptosis / Two-hybrid / ファーウエスタン / 遺伝子ファミリー / ゲノムDNA / SART 1 / 核内蛋白 / ロイシン・ジッパー・モチーフ / ファー・ウェスタン |
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| Research Abstract |
Molecules involved in regulation of cellular proliferation at the G2/M phase have not yet been fully identified. This study investigated the involvement of a recently identified SART-1ィイD2800ィエD2. Protein demonstrated an ability to bind to DNA, and accumulated in the nucleus when the cells were arrested at G2/M phase.
(l) The localization of SART-1 protein was suggested to be in nucleus of eukaryote cells including monkey kidney EBV-transformed Cos 7 cell and human esophageal cancer TE9 cell when the fusion gene with GFP was transfected and observed by conforcal laser microscopy.
(2) SART-1ィイD2800ィエD2 protein is suggested to be a DNA-binding protein by the affinity chromatography of cell lysates of the 293T cells transfected with the HAN/SART-1ィイD2800ィエD2 gene.
(3) Transfection of SART-1 gene induced the G2/M-arrest and suppressed the cell growth of all the cell tested. Furthermore, DNA ladder formation was observed in the SART-1 transfected COS-7 and human cancer cells. The transfection of SART-1 up-regulated the nuclear expression of cyclin B and cdc2, well known components of maturation-promoting factor.
(4) Several candidate genes which encode proteins that had interacted with SART-1 protein were isolated by two-hybrid system.
(5) Three different SART-1 probe protein (N-terminal, middle and C-terminal regions) for far-western blot analysis of SART-1 binding protein candidate obtained by two hybrid system were prepared.
(6) We found that murine SART-1 genomic DNA suggested to be consist of 20 exsons. The sequence of the gene will submit to GenBank.
(7) SART-1 gene was suggested to be expressed in early development of mouse embryo (l4 days) by northern blot analysis.
(8) Several family genes of SART-1 were suggested by northern blot analysis. One of the gene with 3.3kb consist of approximately l488 bp of 5'-region was identical to SART-1 and encoded a protein with 483 amino acid.
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