| Project/Area Number |
06672167
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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 |
Biological pharmacy
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| Research Institution | Chiba University |
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Principal Investigator |
HIROSE Seiyu CHIBA UNIVERSITY,PHARMACEUTICAL SCIENCES,PROFESSOR., 薬学部, 教授 (40009163)
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| Co-Investigator(Kenkyū-buntansha) |
KAKEGAWA Tomohito CHIBA UNIVERSITY,PHARMACEUTICAL SCIENCES,LECTURER., 薬学部, 講師 (80169391)
KUMAGAI Hiroshi CHIBA UNIVERSITY,PADIO ISOTOPE RESEARCH AND EDUCATION CENTER,ASSISTANT PROFESSOR, アイソトープ総合センター, 助教授 (40092051)
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| Project Period (FY) |
1994 – 1995
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| Project Status |
Completed (Fiscal Year 1995)
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| Budget Amount *help |
¥2,100,000 (Direct Cost: ¥2,100,000)
Fiscal Year 1995: ¥600,000 (Direct Cost: ¥600,000)
Fiscal Year 1994: ¥1,500,000 (Direct Cost: ¥1,500,000)
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| Keywords | RAT LIVER / RIBONUCLEASE / PDG-A / AORTIC SMC / 5'-UTR / uORF / mRNA STABILITY / SUBSTRATE SPECIFICITY / 5′非翻訳領域 / 核酸結合タンパク質 / mRNA分解 / 分子生物学 |
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| Research Abstract |
PURIFICATION OF RIBONUCLEASES (HIROSHI KUMAGAI). The novel RNase were purified from rat liver cytosomal fraction ; molecular weight 13 kDa and 16 kDa RNases were termed RNase 13 and RNase 16, respectively. RNase 16 hydrolyzed in the order of poly (C) >yeast total RNA >poly (U) and may be inbolved amember of pancreatic RNase family. On the other hand, RNase 13 hyodrolyzed in the order of poly (U) >yeast total RNA >Poly (C) and may be inbolved a member of plasma RNase family.
POSSIBLE RELATIONSHIP OF THE STABILTY OF mRNA AND ACTIVITY OF A SITE SPECIFIC RNase. (TOMOHITO KAKEGAA). Platelet derived growth factor (PDGF) A plays an important role in the proliferation and chemostasis of animal cells. The most suitable candidate of region of PDGF-A mRNA for site specific degradation was investigated in rat cultured smooth muscle cells. Ultra violet derived crosslinking reaction and RNase protection assay revealed the terminal codon area of the upper open reading frame existing in the 5' untranslated region (5'-UTR) PDGF-A had high potential for the start site of this mRNA.Four kinds of purified rat liver RNases except exo-RNase that was purified from rat liver microsomal fraction did not degrade site specifically with PDGF-A 5'-UTR.This results suggest that site specific degradation of PDGF-A 5'-UTR by three endo-RNases requires additional unknown endogenous factors. The new application of the North-western blot for detection of RNase activity revealed that cytosomal fraction contained many logical candidates for the sequence specific RNase activity. This result suggest that the application of North-western blot for the detection of RNase activity may be useful for determination and isolation of site specific RNA degrading activity.
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