文章基本信息

标题：Induction of p16/INK4a Gene Expression and Cellular Senescence by Toyocamycin
本地全文：下载
作者：Yosuke Kurihara ; Kiyoshi Egawa ; Setsuko Kunimoto 等
期刊名称：Biological and Pharmaceutical Bulletin
印刷版ISSN：0918-6158
电子版ISSN：1347-5215
出版年度：2002
卷号：25
期号：10
页码：1272-1276
DOI：10.1248/bpb.25.1272
出版社：The Pharmaceutical Society of Japan
摘要：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.
关键词：p16 gene;screening;toyocamycin;Sp1