Bi Xin, Yu Qun, Sandmeier Joseph J, Elizondo Susan
Department of Biology, University of Rochester, Rochester, NY 14627, USA.
J Mol Biol. 2004 Dec 3;344(4):893-905. doi: 10.1016/j.jmb.2004.10.002.
Increasing evidence indicates that transcriptionally silent chromatin structure is dynamic and may change its conformation in response to external or internal stimuli. We show that growth temperature affects all three forms of transcriptional silencing in Saccharomyces cerevisiae. In general, increasing the temperature within the range of 23-37 degrees C strengthens HM and telomeric silencing but reduces rDNA silencing. High temperature (37 degrees C) can suppress the silencing defects of histone H4 mutants. We demonstrate that DNA at the silent HML locus becomes more and more negatively supercoiled as temperature increases in a Sir-dependent manner, which is indicative of enhanced silent chromatin. This enhancement of silent chromatin is not dependent on silencers and therefore does not require de novo assembly of silent chromatin. We also present evidence suggesting that MAP kinase-mediated Sir3p hyperphosphorylation, which plays a role in regulating silencing in response to certain stress conditions, is not involved in high temperature-induced strengthening of silencing. In addition, Pnc1p, a positive regulator of Sir2p activity, plays no role in thermal regulation of silencing. Therefore, growth temperature regulates transcriptional silencing by a novel mechanism.
越来越多的证据表明,转录沉默的染色质结构是动态的,可能会响应外部或内部刺激而改变其构象。我们发现生长温度会影响酿酒酵母中所有三种形式的转录沉默。一般来说,在23至37摄氏度范围内升高温度会增强HM和端粒沉默,但会降低rDNA沉默。高温(37摄氏度)可以抑制组蛋白H4突变体的沉默缺陷。我们证明,随着温度升高,沉默的HML位点处的DNA以Sir依赖的方式变得越来越负超螺旋,这表明沉默染色质增强。这种沉默染色质的增强不依赖于沉默子,因此不需要沉默染色质的从头组装。我们还提供了证据表明,在响应某些应激条件下调节沉默中起作用的MAP激酶介导的Sir3p过度磷酸化不参与高温诱导的沉默增强。此外,Sir2p活性的正调节因子Pnc1p在沉默的热调节中不起作用。因此,生长温度通过一种新机制调节转录沉默。