Induction of p16/INK4a Gene Expression and Cellular Senescence by Toyocamycin(Biochemistry/Molecular Biology)

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概要

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• We constructed an assay system of a luciferase reporter with p16/INK4a gene transcriptional regulatory domain to identify p16-inducing substances, and found toyocamycin to induce gene expression from the screening of culture fluids of Streptomyces. Toyocamycin is a nucleoside analog, and it increased the p16 mRNA level in human normal fibroblasts or synovial cells as assessed by Northern blot hybridization or real time RT-PCR. It also induced cellular senescence in normal human fibroblasts. The transcriptional regulatory regions of human p16 gene that were responsible for the induction were analyzed using deletion mutants of the transcriptional regulatory region of p16 linked to the luciferase gene. The DNA fragment -111 to +1 bp from the cap site was sufficient to drive toyocamycin-activated transcription of p16/luciferase reporter. Nucleotide sequences within this domain contained the Sp1- and Ets-binding sequences. Mutations were introduced into these sequences, and the Sp1 sequence was found to be critical for the induction, and this notion was confirmed from gel-mobility shift assay.

• 公益社団法人日本薬学会の論文
• 2002-10-01

著者

• TAKEUCHI Tomio

  Institute of Microbial Chemistry

• Kurihara Youji

  School of Pharmaceutical Sciences, Kitasato University

• Kondo S

  Institute Of Microbial Chemistry

• Takeuchi T

  Bioresource Laboratories Mercian Corp.

• Takeuchi T

  Institute Of Microbial Chemistry

• Kurihara Y

  School Of Pharmaceutical Sciences Kitasato University

• KUNIMOTO Setsuko

  Institute of Microbial Chemistry

• NOSE Kiyoshi

  Department of Microbiology, Showa University School of Pharmacy

• EGAWA Kiyoshi

  Department of Microbiology, Showa University of Pharmaceutical Sciences

• Nose K

  Department Of Microbiology Showa University Of Pharmaceutical Sciences

• Nose K

  Showa Univ. Pharmaceutical Sci. Tokyo Jpn

• Nose Kiyoshi

  Department Of Microbioilogy Showa University School Of Pharmaceutical Sciences

• KUNIMOTO Setsuko

  Numazu Bio-Medical Research Institute, Microbial Chemistry Research Center

• KURIHARA Yosuke

  Department of Microbiology, Showa University School of Pharmaceutical Sciences

• TAKIZAWA Toshio

  Teikyo University Institute of Medical Mycology

• Takeuchi Toshio

  Research And Development Department Kibun Food Chemifa Co. Ltd.

• Nose Kiyoshi

  Department Of Microbiology Showa University Of Pharmaceutical Sciences

• Egawa K

  Department Of Microbiology Showa University Of Pharmaceutical Sciences

• Egawa Kiyoshi

  Department Of Microbiology Showa University Of Pharmaceutical Sciences

• Takeuchi T

  M.c.r.f. Shizuoka Jpn

• Kunimoto S

  Numazu Bio-medical Research Institute Microbial Chemistry Research Center

• Takeuchi Tomio

  Institute For Chemotherapy M.c.r.f.

• Takeuchi T

  Inst. Microbial Chemistry Shinagawa‐ku Jpn

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