转录延伸因子FoTFIIS乙酰化修饰调控尖孢镰刀菌致病性的分子机制

由尖孢镰刀菌侵染植物（120多种）引起的枯萎病是一种世界性的重要病害，对农业生产及人类健康造成严重威胁。前期研究中我们发现尖孢镰刀菌转录延伸因子FoTFIIS第76位赖氨酸存在乙酰化修饰，该位点的突变改变了FoTFIIS的亚细胞定位，导致病原菌侵染番茄的能力显著降低，但机制并不清楚。在此基础上本项目拟通过酵母双杂交文库筛选、基因敲除研究乙酰化修饰调控FoTFIIS亚细胞定位的机理；通过转录组及定量PCR方法分析FoTFIIS调节尖孢镰刀菌致病性的机制；通过基因敲除、Western blot等方法鉴定作用于FoTFIIS的乙酰基转移酶及去乙酰化酶，测定不同生理时期FoTFIIS的乙酰化水平、乙酰基转移酶及去乙酰化酶的表达水平，明确该修饰的动态调控机制。本研究的结果将有助于揭示蛋白乙酰化修饰在病原菌侵染过程中的作用，获得新的药物靶标，为通过设计药物控制枯萎病的发生以及提高作物产量奠定分子基础。

Wilt disease resulting from infection of more than 120 plants by Fusarium oxysporum is an important disease worldwide because of its huge impact on agricultural production and human health. In our previous studies, we found that the transcriptional elongation factor FoTFIIS is acetylated on the lysine residue 76. Mutagenesis of K76 led to altered localization of FoTFIIS and dramatically decreased virulence of F. oxysporum on tomato, but the mechanism remains unclear. In this application, we intend to explore the regulatory mechanism of FoTFIIS localization by acetylation through yeast two-hybrid screening and gene knockout. We will also investigate how FoTFIIS modulates virulence of F. oxysporum through RNA-seq and quantitative reverse transcription PCR analyses. Moreover, we will uncover the dynamic mechanism of FoTFIIS acetylation by identifying the corresponding lysine acetyltransferases and deacetylases acting on FoTFIIS and determining the acetylation level of FoTFIIS and the expression level of these enzymes at different physiological phases through gene knockout, Western blot and so on. This study will not only greatly enhance our understanding of the biological role of lysine acetylation in fungal virulence and to obtain new drug targets, but also lay foundation for controlling plant wilt diseases and improving crop yields by development of small molecules.