| Project/Area Number |
12470501
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| Research Category |
Grant-in-Aid for Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Biological pharmacy
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| Research Institution | HIROSHIMA UNIVERSITY |
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Principal Investigator |
IDE Toshinori Hiroshima University, Faculty of Medicine, Professor, 医学部, 教授 (60012746)
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| Project Period (FY) |
2000 – 2001
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| Project Status |
Completed (Fiscal Year 2001)
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| Budget Amount *help |
¥13,900,000 (Direct Cost: ¥13,900,000)
Fiscal Year 2001: ¥4,000,000 (Direct Cost: ¥4,000,000)
Fiscal Year 2000: ¥9,900,000 (Direct Cost: ¥9,900,000)
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| Keywords | telomere / telomere DNA / extra chromosomal telomere repeats / ECTR / telomere-binding proteins / telomerase / immortalization of cell / proliferative life span |
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| Research Abstract |
Telomere is specialized structure at the end of eukaryotic chromosomes which functions in chromosome stability, positioning, replication and meiosis. Telomere is not composed of common nucleosomal structure but of a complex with several types of telomere specific proteins. Absence of adequate techniques to separate telomeres from non-telormeric parts of chromosome fails to clarify precise structure of telomeres. We found that telomerase-negative immortal human cells contained extra-chromosomal telomere repeats (ECTR) and ECTR was complexed with one of telomere-binding protein, TRF-1. We tried here separation of ECTR protein complex from the bulk chromosome and analysis of protein components in this complex. KMST-6, a human cells line that contains a large number of ECTR, was collected at mitotic phase of the cell cycle and lysed in the buffer containing mild detergent. ECTR-protein complex was separated by differential centrifugation and density gradient centrifugation. When 1 % of total DNA was recovered in ECTR-protein complex fraction, this fraction contains 10 % of total telomere sequence and 1.5 % of total Alu sequence. Further purification by separating nucleosomal chromatin using anti-histone antibody was failed because antibodies were not good for immunoadsorption.
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