| Project/Area Number |
61570122
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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 |
General medical chemistry
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| Research Institution | Shiga University of Medical Science |
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Principal Investigator |
UEDA Kiyoshi Shiga University of Medical Science, 医学部, 教授 (60079704)
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| Co-Investigator(Kenkyū-buntansha) |
OKABE Hidetoshi Shiga University of Medical Science, 医学部, 講師 (70079713)
YOSHIOKA Kuninori Shiga University of Medical Science, 医学部, 助手 (00192419)
UEYAMA Hisao Shiga University of Medical Science, 医学部, 助教授 (30127013)
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| Project Period (FY) |
1986 – 1988
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| Project Status |
Completed (Fiscal Year 1988)
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| Budget Amount *help |
¥2,100,000 (Direct Cost: ¥2,100,000)
Fiscal Year 1988: ¥300,000 (Direct Cost: ¥300,000)
Fiscal Year 1987: ¥400,000 (Direct Cost: ¥400,000)
Fiscal Year 1986: ¥1,400,000 (Direct Cost: ¥1,400,000)
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| Keywords | Nuclear skeleton / Nuclear matrix / Actin / Anti-actin antibody / Western blot / Sarcoma 180 cells / L5178Y cells / cDNA / Phalloidin / サイトカラシン / ウエスタンブロット / サルコーマ180 / L5178Y / メラノーマB16 |
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| Research Abstract |
1. The reactivity of three kinds of anti-actin antibodies with various kinds of actins were examined. The anti-enteric smooth muscle actin antibody reacted only with -cytoplasmic, enteric and aortic smooth muscle actins, while the anti-skeletal muscle actin serum reacted only with skeletal, cardiac and aortic smooth muscle actins. The monoclonal anti-actin antibody reacted with all types of actins. Based on these findings, acidic actins of mouse sarcoma 180 cells, of mouse melanoma B16 cells and of human cytochalasin-resistant KB cells were suggested to be related to -cytoplasmic actin.
2. In vitro translated products of polyARNA from sarcoma 180 cells and from L5178Y cells were analyzed on two-dimensional gels. The acidic actin of sarcoma 180 cells was found to have its own mRNA, while that of L5178Y cells was suggested to be a post-translationally modified form of usual actins.
3. An attempt to separate , and acidic actins without using SDS (hydroxyapatite, chromatofocusing, HPLC on ion-exchange resin・etc.)resultedin a poor success.
4. A cDNA library was constructed from polyARNA of sarcoma 180 cells, and full length cDNA clones of -and -actins were obtained. However, cDNA clones of the acidic actin are not identified yet.
5. In vitro RNA transport system using isolated nuclei of L5178Y cells was developed; phalloidin had no effect of the RNA transport.
6. Cytochalasin had almost no effect on the DNA synthesis of malignant tumor cells, while it caused 80% decrease of DNA synthesis of benign tumor and normal cells.
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