2007 Fiscal Year Final Research Report Summary
Subcellular localization and control of activity of telomerase
| Project/Area Number |
17390091
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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 |
Pathological medical chemistry
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| Research Institution | Kanazawa University |
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Principal Investigator |
MURAKAMI Seishi Kanazawa University, Cancer Research Institute, Emeritus Professor (90019878)
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| Co-Investigator(Kenkyū-buntansha) |
YOSHIOKA Katsuji Kanazawa University, Cancer Research Institute, Professor (60200937)
HONDA Masao Kanazawa University, Graduate Course of Medical Science, Professor (00272980)
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| Project Period (FY) |
2005 – 2007
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| Keywords | Cancer / Virus / Protein / Development and Differentiation / Sienal transduction / Telomerase / Nucleolin / Subcellular localization |
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| Research Abstract |
Since telomerase activity is barely detectable in primary cells but clearly detected in cancerous cells, telomerase has been considered to be a strong candidate of the targets to cancer treatment In the fiscal year, we concentrated in several experiments including purification and characterization of recombinant human telomerase complex, and biological function of the specific interaction of nucleolin and hTERT recovered from a HeLa cell line stably expressing FLAG-hTERT (Murakami), analyses of human primary cells immortalized by hTERT (Honda), and the tyrosine kinases of receptor type which play critical odes in cell proliferation and differentiation (Yoshioka). The followings are the main conclusions of the experiments.
1) The partially purified active telomerase complexes comprise two different complexities, complex I (around 680 kDa), and complex II (around 400 kDa), that are quite alike to those that can be recovered firm the insect cells co-expressing FLAG-hTERT and hTERC. The com
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plex I does not contain Hsp90 and is resistant to Hsp90 inhibitors, whereas complex II contains Hsp90 and is sensitive to Hsp90 inhibitors. Human TEAT in the complex II is rather unstable during incubation in vim and in ring. Such unstable property was not observed with the complex I. Since MG132, a specific proteasome inhibitor, but not MG133, a negative control, stabilized hTERT in the complex Z strongly suggesting that hTERT in the complex If is under degradation pathway under the control of proteasome. The two different complexities of human telomerase seem to imply two different entities of active telomerase in different subcellular localization or its different functions. 2) We have established a cell line of the human primary fibroblasts(BJ cells) stably expressing hTERT, and we compared expression profiles of RI cells and the KJ cells immortalized by MERE Some genes involving in cell cycle progression and cell proliferation were evidently expressed higher in the immortalized 131-hTERT than these in in the primary cells. 3) Nucleolin, one of the specific interacting partners of hTERT can bind to Hepatitis C Virus (HCV) NS5B, RNA-dependent RNA replication of HCV. We evaluated the role of the specific interaction of nucleolin and NS5B in HCV replication. The HCV subreplicon system has been applied for the address We found that the HCV subreplicons harboring alanine substitution mutations or mutation at the residue(s) either one of two critical sequences within NS5B both of which are necessary for the nucleolin-binding could not replicate at all in a transient HCV replication system in HepG2 ml/s., although the wild replicon could support efficient HCV. By introducing transiently RNAi of nucleolin, that down regulated expression of nucleolin by half to one third, reduced HCV evidently reduced HCV replication using the HCV subreplicon system. Taken together, the specific interaction of nucleolin and NS5B is critical for HCV replication. Less
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[Journal Article] Temperature-sensitive defects of the GSPlgene, yeast Ran homologue, activate the Tell-dependent pathway2007
Author(s)
Hayashi, N., Tsurusaki, S., Nagaura, Z., Oki, M., Nishitani, H., Kobayashi, M., Shimizu, H., Yamamoto, K., Nishimoto, T
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Journal Title
Biochem Biophys Res Commun 353(2)
Pages: 330-336
Description
「研究成果報告書概要(欧文)」より
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[Journal Article] Human telomerase exists in two distinct active complexes in vivo2007
Author(s)
Mizuno, H., Khurts, S., Seki, T., Hirota, Y., Kaneko, S., *Murakamk, S
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Journal Title
J. Biochem (Tokyo) 141(5)
Pages: 641-652
Description
「研究成果報告書概要(欧文)」より
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[Journal Article] Hepatitis B Virus X protein overcomes oncogenic RAS-induced senescence in human immortalized cells2007
Author(s)
Oishi, N., Khurts, S., Nakamoto, Y., Honda, M., Kaneko, S., *Murakami, S
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Journal Title
Cancer Science 98(10)
Pages: 1540-1548
Description
「研究成果報告書概要(欧文)」より
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[Journal Article] Mutations in Ran system affected telomere silencing in Saccharomyces cerevisiae2007
Author(s)
Hayashi, N., Kobayashi, M., Shimizu, H., Yamamoto, K., Murakami, C., Nishimoto, T
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Journal Title
Biochem Biophys Res Commun 363(3)
Pages: 788-794
Description
「研究成果報告書概要(欧文)」より
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[Journal Article] Regulation of N-cadherin-based cell-cell interaction by JSAPI scaffold in PC12h cells2007
Author(s)
Bayarsaikhan, M., Takino, T., Gantulga, D., Sato, H., Ito, T., Yoshioka, K
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Journal Title
Biochem Biophys Res Commun 353(2)
Pages: 357-362
Description
「研究成果報告書概要(欧文)」より
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[Journal Article] Matsuzawa-Nagata N, Uchikata M, Nakamura S, Matoba R, Tanino M, Matsubara K, Kaneko S. Gene expression profiles in peripheral blood mononuclear cells reflect the pathophysiology of type 2 diabetes2007
Author(s)
Takamura, T., Honda, M., Sakai, Y., Ando, H., Shimizu, A., Ota, T., Sakurai, M., Misu, H., Kurita, S
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Journal Title
Biochem Biophys Res Commun 361(2)
Pages: 379-384
Description
「研究成果報告書概要(欧文)」より
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[Presentation] Increased dyidative stress precodes the onset of high-fat diet-induced insulin resistance and obesity2008
Author(s)
Takamura, T., Nagata, N., Komura, T., Sakai, Y., Honda, M., Kaneko, S
Organizer
Keystone Symposia(X3)
Year and Date
2008-02-22
Description
「研究成果報告書概要(欧文)」より
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[Presentation] Different Signaling Pathways in the Liver of Patients with Chronic Hepatitis Induced by Hepatitis B and Hepatitis C Virus Infectionn2007
Author(s)
Honda, M., Yamashita, T., Ueda, T., Takatori, H., Nishino, R., Kaneko, S
Organizer
APASL
Place of Presentation
Kyoto
Year and Date
20070000
Description
「研究成果報告書概要(欧文)」より
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[Presentation] Comprehensive gene expression analysis of iron metabolism-related genes in patients with chronic viral hepatitis and hepatocellular carcinoma2007
Author(s)
Honda, M., Ueda, T., Yamashita, T., Nishino, R., Takatori, H., Kaneko, S
Organizer
58th American Association of Hepatology
Year and Date
2007-11-06
Description
「研究成果報告書概要(欧文)」より
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[Presentation] MicroRNA expression profiling combined with analysis of target gene expression in chronic viral hepatitis and hepatocellular carcinoma2007
Author(s)
Ura, S., Honda, M., Ueda, T., Nishino, R., Takatori, H., Nakamura, M., Kaneko, S
Organizer
58th American Association of Hepatology
Year and Date
2007-11-03
Description
「研究成果報告書概要(欧文)」より
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[Presentation] Functional interaction of hepatitis C virus NS5B with nucleolin GAR domain
Author(s)
Shimakami, T., Kusakawa, T., Honda, M., Kaneko, S., Yohioka, K., Murakami, S
Organizer
15th International symposium on Hepatitis C Virus & Related Viruses
Place of Presentation
Texas
Year and Date
00001005-09
Description
「研究成果報告書概要(欧文)」より
