| Project/Area Number |
09670139
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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 |
General medical chemistry
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| Research Institution | Kyoto Pharmaceutical University |
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Principal Investigator |
HATAYAMA Takumi Kyoto pharmaceutical University, Professor, 薬学部, 教授 (10094484)
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| Co-Investigator(Kenkyū-buntansha) |
YAMAGISHI Nobuyuki Kyoto pharmaceutical University, research associate, 薬学部, 助手 (60298685)
YASUDA Kunihiko Kyoto pharmaceutical University, research associate, 薬学部, 助手 (50278446)
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| Project Period (FY) |
1997 – 1998
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| Project Status |
Completed (Fiscal Year 1998)
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| Budget Amount *help |
¥2,600,000 (Direct Cost: ¥2,600,000)
Fiscal Year 1998: ¥800,000 (Direct Cost: ¥800,000)
Fiscal Year 1997: ¥1,800,000 (Direct Cost: ¥1,800,000)
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| Keywords | Stress protein / HSP105 / HSP105 gene / Apoptosis / Embryo developement / Heat shock protein / Mouse |
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| Research Abstract |
1. The 105-kDa heat shock protein (HSP105) is a member of the high molecular mass heat shock protein family. To elucidate the genomic structure of mouse HSP105 and to examine the regulation of expression of its gene, we have isolated and characterized the mouse HSP105 gene including about 1.2kb of the 5'-flanking region. (1) The mouse HSP105 gene spans about 22 kb, consisting of 18 exons separated by 17 introns. (2) Southern blotting analysis revealed the existence of a single copy of HSP105. (3) Primer extension analysis revealed that the transcription initiation site was located 165 bp upstream of the ATG translation initiation codon. (4) The 5'-promoter region of the HSP105 gene contained a TATA box, a CAAT box, an inverted CAAT box and two GC boxes. Two heat shock element (HSE) sequences were found as four nGAAn repeats at nt -64 and nt -128. (5) Promoter analysis using deletion derivatives revealed that a minimal region which contained the two consensus HSE sequences was active in response to heat shock and also for constitutive expression of the gene.
2. To examine the role of HSP105 for differentiation and apoptosis.of mouse teratocarcinoma F9 cells, mouse HSP105alpha cDNA expression plasmid constructed with pcDNA3 vector was introduced into F9 cells, and isolated two HSP105alpha-overexpressing cells which expressed HSP105 at 2-3-fold higher levels than parent cells. (2) In response to various stresses such as heat shock, actinomycin D, etoposide and hydrogen peroxide, the HSP105alpha-overexpressing cells were more sensitive than parent cells or the cells transfected with pcDNA3 vector. (3) Since the increased sensitivity to these stresses was due to an increase of apoptotic cell death, HSP105alpha was suggested to enhance apoptosis. Experiments to elucidate precise mechanisms of HSP105alpha for apoptosis are now in progress.
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