Fermi Beatrice, Bosio Maria Cristina, Dieci Giorgio
Dipartimento di Bioscienze, Università degli Studi di Parma, Parco Area delle Scienze 23/A, 43124 Parma, Italy.
Dipartimento di Bioscienze, Università degli Studi di Parma, Parco Area delle Scienze 23/A, 43124 Parma, Italy
Nucleic Acids Res. 2016 Jul 27;44(13):6113-26. doi: 10.1093/nar/gkw194. Epub 2016 Mar 25.
In Saccharomyces cerevisiae, ribosomal protein gene (RPG) promoters display binding sites for either Rap1 or Abf1 transcription factors. Unlike Rap1-associated promoters, the small cohort of Abf1-dependent RPGs (Abf1-RPGs) has not been extensively investigated. We show that RPL3, RPL4B, RPP1A, RPS22B and RPS28A/B share a common promoter architecture, with an Abf1 site upstream of a conserved element matching the sequence recognized by Fhl1, a transcription factor which together with Ifh1 orchestrates Rap1-associated RPG regulation. Abf1 and Fhl1 promoter association was confirmed by ChIP and/or gel retardation assays. Mutational analysis revealed a more severe requirement of Abf1 than Fhl1 binding sites for RPG transcription. In the case of RPS22B an unusual Tbf1 binding site promoted both RPS22B and intron-hosted SNR44 expression. Abf1-RPG down-regulation upon TOR pathway inhibition was much attenuated at defective mutant promoters unable to bind Abf1. TORC1 inactivation caused the expected reduction of Ifh1 occupancy at RPS22B and RPL3 promoters, but unexpectedly it entailed largely increased Abf1 association with Abf1-RPG promoters. We present evidence that Abf1 recruitment upon nutritional stress, also observed for representative ribosome biogenesis genes, favours RPG transcriptional rescue upon nutrient replenishment, thus pointing to nutrient-regulated Abf1 dynamics at promoters as a novel mechanism in ribosome biogenesis control.
在酿酒酵母中,核糖体蛋白基因(RPG)启动子显示出Rap1或Abf1转录因子的结合位点。与Rap1相关的启动子不同,一小部分依赖Abf1的RPG(Abf1-RPG)尚未得到广泛研究。我们发现,RPL3、RPL4B、RPP1A、RPS22B和RPS28A/B共享一种共同的启动子结构,在一个保守元件上游有一个Abf1位点,该保守元件与Fhl1识别的序列相匹配,Fhl1是一种转录因子,它与Ifh1一起协调Rap1相关的RPG调控。通过染色质免疫沉淀(ChIP)和/或凝胶阻滞试验证实了Abf1和Fhl1与启动子的结合。突变分析表明,与Fhl1结合位点相比,Abf1结合位点对RPG转录的要求更为严格。就RPS22B而言,一个不寻常的Tbf1结合位点促进了RPS22B和内含子宿主的SNR44表达。在无法结合Abf1的缺陷突变启动子上,TOR途径抑制后Abf1-RPG的下调程度大大减弱。TORC1失活导致Ifh1在RPS22B和RPL3启动子上的占有率如预期那样降低,但出乎意料的是,这导致Abf1与Abf1-RPG启动子的结合大幅增加。我们提供的证据表明,营养应激时Abf1的募集(在代表性的核糖体生物发生基因中也观察到)有利于营养补充后RPG的转录拯救,从而指出启动子处营养调节的Abf1动态是核糖体生物发生控制中的一种新机制。