| Project/Area Number |
02454142
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| Research Category |
Grant-in-Aid for General Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Research Field |
General medical chemistry
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| Research Institution | Chiba University |
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Principal Investigator |
TATIBANA Masamiti Chiba University School of Medicine, Professor, 医学部, 教授 (50009081)
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| Co-Investigator(Kenkyū-buntansha) |
SONODA Tomoko Chiba University School of Medicine, Research Associate, 医学部, 教務職員 (20143307)
ISHIZUKA Toshiharu Chiba University School of Medicine ,Assistant, 医学部, 助手 (50232294)
ISHIJIMA Sumio Chiba University School of Medicine ,Assistant, 医学部, 助手 (70184520)
SUZUKI Nobuo Chiba University School of Medicine ,Assistant Professor, 医学部, 助教授 (90111426)
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| Project Period (FY) |
1990 – 1992
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| Project Status |
Completed (Fiscal Year 1992)
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| Budget Amount *help |
¥6,400,000 (Direct Cost: ¥6,400,000)
Fiscal Year 1992: ¥1,600,000 (Direct Cost: ¥1,600,000)
Fiscal Year 1991: ¥1,700,000 (Direct Cost: ¥1,700,000)
Fiscal Year 1990: ¥3,100,000 (Direct Cost: ¥3,100,000)
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| Keywords | Nucleic acid precursors / Phosphoribosylpyro-phosphate synthetase / Isoform / Feedback inhibition / Promoter region / Magnesium ion / Mitogenic stimuli / Fluorescent dye / 制御タンパク / アイソフォ-ム / フィ-ドバック阻害 / 発現ベクタ- |
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| Research Abstract |
1. Studies on phosphoribosylpyrophosphate (PRPP) synthetase (1) Rat PRPP synthetase subunits I and II (PRS I and PRS II, respectively) were expressed in Escherichia coli. The recombinant isoforms were purified to apparent homogeneity and characterized. The two isoforms differed in sensitivity to inhibition by ADP and GDP, and heat inactivation. (2) cDNA clones for human PRS I were isolated from a glioblastoma cell line MGC 1 cDNA library.The deduced amino acid sequences were identical between human and rat PRS I. Based on the evolutionary rate of amino acid substitution, the PRS I and II genes probably diverged about 760 million years ago. (3) The 5' regions of the human PRS I and PRS II genes were isolated and sequenced. Chloramphenicol acetyltransferase/promoter fusion assays suggest that the 5' flanking regions cloned possess the promoter activities and contribute to the cell-differential expression of these two genes. 2. Studies on changes in cytosolic free Mg^<2+> concentrations ([Mg^<2+>]_i) after mitogenic stimulation in single mouse Swiss 3T3 fibroblasts (1) Mitogen-induced changes in [Mg^<2+>]_i were measured at the single-cell level, using digital ratio imaging microscopy of the fluorescent probe mag-fura-2. (2) Stimulation of the cells with bombesin or epidermal growth factor in combination with insulin led to a significant increase in [Mg^<2+>]_i after 30-60 min. These results provide direct evidence for the mobilization of Mg^<2+> as an early cellular response to growth factors. It is possible that free Mg^<2+>-sensitive enzymes play regulatory roles in signal transduction through changes in [Mg^<2+>]_i.
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