Human telomerase exists in two distinct active complexes in vivo
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概要
- 論文の詳細を見る
Telomerase, a stable complex of telomerase reverse transcriptase (TERT) and template RNA (TERC), is responsible for telomere maintenance. During purification trials of recombinant human telomerase of the two components reconstituted in insect cells, we identified two complexes of human telomerase of molecular masses 680 and 380 kDa, both of which retain telomerase activity in vitro. We show here that the former complex does not include Hsp90 (heat shock protein 90) and its telomerase activity is resistant to Hsp90 inhibitors, whereas the latter contains Hsp90 and its telomerase activity is sensitive to Hsp90 inhibitors. N-terminal of FLAG-hTERT in the former is exposed, as this complex was efficiently purified with anti-FLAG M2 affinity resin. We also identified two different telomerase complexes in HeLa cells, in addition to ectopically expressed hTERT. Most of endogenous hTERT and FLAG-hTERT was detected around 680 kDa. These two complexes in HeLa cells have the same properties as their respective reconstituted telomerases. The unstable property of the telomerase complex with Hsp90, especially in the presence of Hsp90 inhibitors, was due to proteasome-mediated degradation of hTERT, since proteasome inhibitors prevented hTERT degradation in vivo. To our knowledge, this is the first demonstration of two distinct active complexes of human telomerase ectopically expressed in insect and mammalian cells. © 2007 The Japanese Biochemical Society.
- 日本生化学会 = Japanese Biochemical Societyの論文
- 2007-05-01
著者
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Kaneko Shuichi
Department Of Disease Control And Homeostasis Kanazawa University Graduate School Of Medical Science
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MURAKAMI Seishi
Department of Signal Transduction, Cancer Research Institute, Graduate School of Medicine, Kanazawa
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Khurts Shilagardi
金沢大学がん研究所 細胞機能統御研究分野
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Khurts Shilagardi
Department Of Molecular Oncology Cancer Research Institute Kanazawa University
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MIZUNO Hideki
Department of Molecular Oncology, Cancer Research Institute, Kanazawa University
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SEKI Takahiko
New Product Research Laboratories III R&D Division, Daiichi Pharmaceutical Company Ltd.
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HIROTA Yasuhide
New Product Research Laboratories III R&D Division, Daiichi Pharmaceutical Company Ltd.
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Mizuno Hideki
Department Of Microbiology Dokkyo University School Of Medicine
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Mizuno Hideki
Department Of Molecular Oncology Cancer Research Institute Kanazawa University
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Seki Takahiko
New Product Research Laboratories Iii R&d Division Daiichi Pharmaceutical Company Ltd.
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Kaneko Shuichi
Department Of Cancer Gene Regulation Kanazawa University
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Kaneko Shuichi
Department Of Gastroenterology Graduate School Of Medical Science Kanazawa University
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Murakami Seishi
Department Of Molecular Oncology Cancer Research Institute Kanazawa University
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Hirota Yasuhide
New Product Research Laboratories Iii R&d Division Daiichi Pharmaceutical Company Ltd.
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