| Project/Area Number |
11557130
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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 | 展開研究 |
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| Research Field |
Morphological basic dentistry
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| Research Institution | Hokkaido University |
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Principal Investigator |
SHINDOH Masanobu Hokkaido Univ. Grad. School of Dent. Med. Asso. Prof., 大学院・歯学研究科, 助教授 (20162802)
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| Co-Investigator(Kenkyū-buntansha) |
KOBAYASHI Masanobu Hokkaido Univ. Grad. School of Dent. Med. Asso. Prof., 遺伝子病制御研究所, 助教授 (80241321)
YASUDA Motoaki Hokkaido Univ. Grad. School of Dent. Med. Asso. Prof., 大学院・歯学研究科, 助教授 (90239765)
HIGASHINO Fumihiro Hokkaido Univ. Grad. School of Dent. Med. Inst., 大学院・歯学研究科, 助手 (50301891)
澤田 幸治 札幌医科大学, 医学部・附属がん研究所, 助教授 (80111128)
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| Project Period (FY) |
1999 – 2000
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| Project Status |
Completed (Fiscal Year 2001)
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| Budget Amount *help |
¥13,000,000 (Direct Cost: ¥13,000,000)
Fiscal Year 2000: ¥6,900,000 (Direct Cost: ¥6,900,000)
Fiscal Year 1999: ¥6,100,000 (Direct Cost: ¥6,100,000)
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| Keywords | E1AF / p53 / E4orf6 / ウイルスベクター / E1AFプロモーター / アポトーシス / アデノウィルスベクター / 転写調節 |
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| Research Abstract |
ElAF and Bax activation was investigated since ElAF was shown to activate promoter activity of p21waf1/cip1, a cell cycle inhibitory molecule.
CAT (Chloramphenicol acetyl transferase) assay was carried out to determine the promoter activity of bax by transfectipn with bax reporter plasmid, ElAF, wild type (wt) or mutant (mt) p53 expression plasmids into a p53 diffident cell line. CAT activity increased with ElAF in a dose-dependent manner and synergistical increase of the promoter activity was observed with wt p53 and ElAF. Protein expression of Bax was confirmed in ElAF transfected cells by Western blotting. Growth suppression assay was performed and decreased colony formation was observed in ElAF transfected p53 diffident cells. These results suggest that ElAF associates with p53 to induce apoptosis through activating bax.
E4orf6 is a newly identified viral oncoprotein of the adenovirus. E4orf6 markedly enhances the ability of transformed rat BRK and human 293 cells to form tumors in nude mice. The E4orf6 protein has been shown to interact with p53 and p73 gene products and to block the induction of apoptosis which is mediated by these antioncogenes. We investigated whether or not E4orf6 can influence molecules that induce apoptosis other than the p53 family, because p53-independent apoptosis pathways have also been proposed. We demonstrate that the adenovirus E4orf6 protein suppresses the apoptotic activity of BNIP3, a member of a group of an apoptosis initiators of the Bcl-2 family. Expression of E4orf6 inhibits BNIP3-mediated apoptosis and its mitochondrial translocation. E4orl6 interacts with BNIP3. In addition, the nuclear export signal of E4orf6 is important for these functions of E4orf6. Our data suggest that the pro-apoptotic protein of the Bcl-2 family is a target of the E4orf6 protein in transformed cells.
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