| Project/Area Number |
05670148
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| Research Category |
Grant-in-Aid for General Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Research Field |
Pathological medical chemistry
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| Research Institution | Kochi Medical School |
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Principal Investigator |
TANIGUCHI Taketoshi Kochi Medical School Medical research Laboratory Associate Professor, 医学部, 助教授 (90127944)
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| Project Period (FY) |
1993 – 1994
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| Project Status |
Completed (Fiscal Year 1994)
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| Budget Amount *help |
¥2,100,000 (Direct Cost: ¥2,100,000)
Fiscal Year 1994: ¥1,000,000 (Direct Cost: ¥1,000,000)
Fiscal Year 1993: ¥1,100,000 (Direct Cost: ¥1,100,000)
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| Keywords | MHC class II / Gene Regulation / Poly (ADP-ribose) / Fusion Protein / Macrophage / B Lymphocyte / MHC class II / ポリADP-リボース / マクロファージ / ポリADPR / HLA-DR / IFNgamma / Reporter gene / 遺伝子調節 / アンチセンスRNA / Promoter |
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| Research Abstract |
We have demonstrated that the high expression of poly (ADP-ribose) synthetase (PARS) caused the depression of MHC class II molecules. In B cell lines, which constitutively express the class II molecule, there are significant amounts of PARS.Thus, we attempted to detect a cell type-specific trans-acting factor which interacted with PARS and regulated the MHC class II gene.
I.We constructed a fusion protein by using the automodification and DNA binding domains of PARS.We labeled nuclear proteins with ^<35>S-Met in B cells and macrophage cells and the ^<35>S-labeled nuclear proteins were incubated with the PARS-fusion protein. We detected a 45kDa and 35kDa binding partner for PARS in B cells.
II.We prepared a probe DNA of human MHC class II genomic DNA in the region of DNase I hypersensitive site II(DHSII). We prepared nuclear proteins from class II-positive cells, class II-inducible cells and negative cells and analyzed them by South-western blotting by the use of the DHS II DNA probe. We detected a 35kDa nuclear protein which interacted with the DNA probe. Whether or not the DHS II-associated 35kDa protein interacts with PARS is still under investigation.
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