González-Lamothe Rocío, Boyle Patrick, Dulude Annie, Roy Vicky, Lezin-Doumbou Cyr, Kaur Gidda Satinder, Bouarab Kamal, Després Charles, Brisson Normand
Department of Biochemistry, Université de Montréal, Montréal, Quebec, Canada H3C 3J7.
Plant Cell. 2008 Nov;20(11):3136-47. doi: 10.1105/tpc.108.061721. Epub 2008 Nov 21.
Transcriptional reprogramming is critical for plant disease resistance responses. In potato (Solanum tuberosum), the marker gene PATHOGENESIS-RELATED-10a (PR-10a) is transcriptionally activated by pathogens, wounding, or elicitor treatment. Activation of PR-10a requires the recruitment of the activator Why1 to its promoter. In addition, PR-10a is negatively regulated by the repressor SEBF (for Silencer Element Binding Factor). Here, we show through a yeast two-hybrid screen that SEBF interacts with Pti4, which has been shown to be a transcriptional activator. SEBF recruits Pti4 via its consensus sequence-type RNA binding domain, while Pti4 is recruited to SEBF by means of its ethylene-response factor domain. In vivo plant transcription assays confirmed that SEBF interacts with Pti4 to form a repressosome, showing that Pti4 can also play a role in transcriptional repression. Chromatin immunoprecipitation revealed that both SEBF and Pti4 are recruited to the PR-10a promoter in uninduced conditions only and that the recruitment of Pti4 is dependent on the presence of SEBF, consistent with the fact that there is no Pti4 consensus binding site in PR-10a. Unexpectedly, we also demonstrated that recruitment of SEBF was dependent on the presence of Pti4, thereby explaining why SEBF, itself a repressor, requires Pti4 for its repressing function.
转录重编程对植物抗病反应至关重要。在马铃薯(Solanum tuberosum)中,标记基因病程相关蛋白10a(PR - 10a)在病原体、创伤或激发子处理下会被转录激活。PR - 10a的激活需要激活因子Why1募集到其启动子上。此外,PR - 10a受到阻遏因子SEBF(沉默子元件结合因子)的负调控。在此,我们通过酵母双杂交筛选表明,SEBF与已被证明是转录激活因子的Pti4相互作用。SEBF通过其共有序列型RNA结合结构域募集Pti4,而Pti4则通过其乙烯反应因子结构域被募集到SEBF上。体内植物转录分析证实,SEBF与Pti4相互作用形成一个阻遏体,表明Pti4也能在转录抑制中发挥作用。染色质免疫沉淀显示,仅在未诱导条件下SEBF和Pti4都被募集到PR - 10a启动子上,并且Pti4的募集依赖于SEBF的存在,这与PR - 10a中不存在Pti4共有结合位点的事实一致。出乎意料的是,我们还证明了SEBF的募集依赖于Pti4的存在,从而解释了为什么作为阻遏因子的SEBF需要Pti4来发挥其抑制功能。