Claypool Jonathan A, French Sarah L, Johzuka Katsuki, Eliason Kristilyn, Vu Loan, Dodd Jonathan A, Beyer Ann L, Nomura Masayasu
Department of Biological Chemistry, University of California, Irvine, Irvine, California 92697-1700, USA.
Mol Biol Cell. 2004 Feb;15(2):946-56. doi: 10.1091/mbc.e03-08-0594. Epub 2003 Oct 31.
Yeast cells entering into stationary phase decrease rRNA synthesis rate by decreasing both the number of active genes and the transcription rate of individual active genes. Using chromatin immunoprecipitation assays, we found that the association of RNA polymerase I with the promoter and the coding region of rDNA is decreased in stationary phase, but association of transcription factor UAF with the promoter is unchanged. Similar changes were also observed when growing cells were treated with rapamycin, which is known to inhibit the Tor signaling system. Rapamycin treatment also caused a decrease in the amount of Rrn3p-polymerase I complex, similar to stationary phase. Because recruitment of Pol I to the rDNA promoter is Rrn3p-dependent as shown in this work, these data suggest that the decrease in the transcription rate of individual active genes in stationary phase is achieved by the Tor signaling system acting at the Rrn3p-dependent polymerase recruitment step. Miller chromatin spreads of cells treated with rapamycin and cells in post-log phase confirm this conclusion and demonstrate that the Tor system does not participate in alteration of the number of active genes observed for cells entering into stationary phase.
进入稳定期的酵母细胞通过减少活性基因数量和单个活性基因的转录速率来降低rRNA合成速率。利用染色质免疫沉淀分析,我们发现RNA聚合酶I与rDNA启动子和编码区的结合在稳定期减少,但转录因子UAF与启动子的结合未改变。当生长中的细胞用雷帕霉素处理时也观察到类似变化,已知雷帕霉素可抑制Tor信号系统。雷帕霉素处理还导致Rrn3p - 聚合酶I复合物的量减少,类似于稳定期。因为如本研究所示,Pol I募集到rDNA启动子是Rrn3p依赖性的,这些数据表明稳定期单个活性基因转录速率的降低是通过Tor信号系统在Rrn3p依赖性聚合酶募集步骤起作用来实现的。雷帕霉素处理的细胞和对数后期细胞的Miller染色质铺展证实了这一结论,并表明Tor系统不参与进入稳定期细胞中观察到的活性基因数量的改变。