2002 Fiscal Year Final Research Report Summary
Role for protein phosphatase type 2A in infiltration of mast cells into tissues
| Project/Area Number |
13670215
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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 |
Experimental pathology
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| Research Institution | Osaka University |
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Principal Investigator |
ITO Akihiko Osaka University Graduate School of Medicine Assistant Professor, 医学系研究科, 助手 (80273647)
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| Project Period (FY) |
2001 – 2002
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| Keywords | MITF / Subtraction / Adhesion molecule / SgIGSF / SynCAM |
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| Research Abstract |
MITF is a basic-helix-loop-helix-leucine zipper-type transcription factor. The mutant mi and Mi^<wh> alleles encode MITFs with deletion and alteration of a single amino acid, respectively, while the tg is a null mutation. When cultured mast cells (CMCs) are established from spleens of C57BL/6 (B6) wild-type (+/+) mice and cocultured on a monolayer ofNIH/3T3 fibroblasts, a considerable number of CMCs adhered to NIH/3T3 fibroblasts. In contrast, the number of CMCs derived from C57BL/6 (B6)-mi/mi, B6-Mi^<wh>/Mi^<wh> and B6-tg/tg mice that adhered to NIH/3T3 fibroblasts was one third that ofB6-+/+ CMCs. We subtracted a cDNA library ofB6-+/+ CMCs with messenger RNAs expressed by B6-mi/mi CMCs and found a clone encoding SgIGSF, a recently identified member of the immunoglobulin superfamily. Northern and Western blot analyses revealed that SgIGSF was expressed in B6-+/+ CMCs but not in CMCs derived from MITF mutants. Immunocytochemical analysis showed that SgIGSF localized to the cell-cell contact areas between B6-+/+ CMCs and NIH/3T3 fibroblasts. Transfection of B6-mi/mi and B6-tg/tg CMCs with SgIGSF cDNA normalized their adhesion to NIH/3T3 fibroblasts. NIH/3T3 fibroblasts did not express SgIGSF, indicating that SgIGSF acts as a heterophilic adhesion molecule. Transfection of B6-tg/tg CMCs with normal MITF cDNA elevated their SgIGSF expression to normal levels. Wild-type MITF, but not mi-type MITF, could bind to a motif in the SgIGSF gene promoter and this association resulted in a significant transactivation of the promoter. These results indicated that SgIGSF mediated the adhesion of CMCs to fibroblasts and that the transcription of SgIGSF was critically regulated by MITF.
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Research Products
(16 results)
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[Publications] Morii E, Ogihara H, Kim DK, Ito A, Oboki K, Lee YM, Jippo T, Nomura S, Maeyama K, Lamoreux ML, and Kitamura Y: "Importance of leucine zipper domain of mi transcription factor (MITF) for differentiation of mast cells demonstrated using mi^<ce>/ mi^<ce> mutant mice of which MITF lacks the zipper domain"Blood. 97. 2038-2044 (2001)
Description
「研究成果報告書概要(欧文)」より
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[Publications] Watabe K, Ito A, Asada H, Endo Y, Kobayashi T, Nakamoto K, Kami S, Takao S, Shinomura Y, Aikou T, Yoshikawa K, Matsuzawa Y, Kitamura Y, and Nojima H: "Structure, expression and chromosome mapping of MLZE, a novel gene which is preferentially expressed in metastatic melanoma cells"Jpn J Cancer Res. 92. 140-151 (2001)
Description
「研究成果報告書概要(欧文)」より
