2002 Fiscal Year Final Research Report Summary
Functional Contribution of Small Maf Proteins in Hematopoietic Cells
| Project/Area Number |
13670112
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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 |
General medical chemistry
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| Research Institution | University of Tsukuba |
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Principal Investigator |
MOTOHASHI Hozumi University of Tsukuba, Institute of Basic Medical Sciences, Lecturer, 基礎医学系, 講師 (00282351)
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| Co-Investigator(Kenkyū-buntansha) |
YAMAMOTO Masayuki University of Tsukuba, Institute of Basic Medical Sciences, Professor, 基礎医学系, 教授 (50166823)
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| Project Period (FY) |
2001 – 2002
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| Keywords | small Maf proteins / megakaryocytes / transgenic mouse / proplatelet formation / transcription / subtraction |
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| Research Abstract |
(1) Transcriptional regulation of mafG gene. Three independent promoters were identified by 5'-RACE in mafG gene. A 7 kbp-genomic fragment containing the mafG promoters were able to direct tissue-specific reporter gene expression in brain, lens, lung and kidney, but not in hematopoietic cells of transgenic mouse. We are going to test additional 3 kbp fragment downstream of mafG gene.
(2) Subtraction analysis of mafG-null megakaryocytes. mRNA derived from proplatelet formation (PPF)-defective mafG-/-::mafK+/- megakaryocytes were subtracted by mRNA of control wild type megakaryocytes. Several clones obtained from the subtraction were found to be related to cytoskeletal proteins, such as actin and tubulin. One novel clone, named #325, was also obtained, and it was found to be conserved among various vertebrates from fish to human. The expression of #325 increased according to the differentiation of immature hematopoietic cells to megakaryocytes. We are generating #325-overexpressing transgenic mouse and disaipting #325 gene in mouse to examine the in vivo function of the gene.
(3) Analysis of MafG C-terminal region. MafG C-terminal region was deleted to generate MafGAC. The mutant molecule also repressed the transcription as well as the wild type MafG when they were overexpressed in cultured cells, When the synergistic transcriptional activity of MafGAC with p45 was compared to that of MafG, MafGAC showed much stronger transcriptional activity than wild type MafG. We are going to test the function of MafGAC in vivo.
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[Publications] Noda, S., Harada, N., Hida, A., Fujii-Kuriyama, Y., Motohashi, H., et al.: "Gene expression of detoxifying enzymes in AhR and Nrt2 compound null mutant mouse"Biochem.Biophys.Res.Commun.. 303. 105-111 (2003)
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[Publications] Yoh, K., Sugawara, T., Motohashi, H., Takahama ,Y, Koyama, A., Yamamoto, M., Takahashi, S.: "Transgenic over-expression of MafK suppresses T cell proliferation and function in vivo"Genes Cells. 6. 1055-1066 (2001)
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[Publications] Kusunoki, H., Motohashi, H., Katsuoka, F., Morohashi, A., Yamamoto, M. and Tanaka, T.: "Solution structure of the DNA-binding domain of MafG"Nature Struct. Biol.. 4. 252-256 (2002)
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[Publications] Kwak, M.K., Wakabayashi, N., Itoh, K., Motohashi,H., Yamamoto, M.., Kensler, T.W.: "Modulation of gene expression by cancer chemopreventive dithiolethiones through the Keap1-Nrf2 pathway : Identification of novel gene clusters for cell survival"J. Bioi. Chem.. 278. 8135-8145 (2003)
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[Publications] Katsuoka, F., Motohashi, H., Tamagawa, Y., Kure, S., Igarashi, K., Engel, J.D., Yamamoto, M.: "Small Maf compound mutants display CNS neuronai degeneration, aberrant transcription and Bach protein misl ocalozati on coinci dent with myoclonus and abnormal startle response"Mol. Cell. Biol.. 23. 1163-1174 (2003)
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「研究成果報告書概要(欧文)」より
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[Publications] Noda, S., Harada, N., Hida, A., Fujii-Kuriyama, Y., Motohashi, H., Yamamoto, M.: "Gene expression of detoxifying enzymes in AhR and Nrf2 compound null mutant mouse"Biochem. Biophys. Res. Commun.. 303. 105-111 (2003)
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「研究成果報告書概要(欧文)」より
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[Publications] Moriguchi, T., Motohashi, H., Hosoya, T, Nakajima, O., Takahashi, S., Ohsako, S., Aoki, Y., Nishimura, N., Tohyama, C., Fujii-Kuriyama, Y., Yamamoto, M.: "Distinct response to dioxin in an arylhydrocarbon receptor (AhR.)-hurnanized mouse"Proc. Natl. Acad. Sci. USA. in press.
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