Lapham Rachelle A, Lee Lan-Ying, Xhako Eder, Gómez Esteban Gañán, Nivya V M, Gelvin Stanton B
Department of Biological Sciences, Purdue University, West Lafayette, IN, United States.
Departamento de Ciencias Biológicas, Universidad EAFIT, Medellín, Colombia.
Front Plant Sci. 2021 Jun 4;12:684192. doi: 10.3389/fpls.2021.684192. eCollection 2021.
effector protein VirE2 is important for plant transformation. VirE2 likely coats transferred DNA (T-DNA) in the plant cell and protects it from degradation. VirE2 localizes to the plant cytoplasm and interacts with several host proteins. Plant-expressed VirE2 can complement a mutant strain to support transformation. We investigated whether VirE2 could facilitate transformation from a nuclear location by affixing to it a strong nuclear localization signal (NLS) sequence. Only cytoplasmic-, but not nuclear-localized, VirE2 could stimulate transformation. To investigate the ways VirE2 supports transformation, we generated transgenic plants containing a gene under the control of an inducible promoter and performed RNA-seq and proteomic analyses before and after induction. Some differentially expressed plant genes were previously known to facilitate transformation. Knockout mutant lines of some other VirE2 differentially expressed genes showed altered transformation phenotypes. Levels of some proteins known to be important for transformation increased in response to VirE2 induction, but prior to or without induction of their corresponding mRNAs. Overexpression of some other genes whose proteins increased after VirE2 induction resulted in increased transformation susceptibility. We conclude that cytoplasmically localized VirE2 modulates both plant RNA and protein levels to facilitate transformation.
效应蛋白VirE2对植物转化很重要。VirE2可能在植物细胞中包裹转移DNA(T-DNA)并保护其不被降解。VirE2定位于植物细胞质并与多种宿主蛋白相互作用。植物表达的VirE2可以互补突变菌株以支持转化。我们研究了VirE2是否可以通过附加一个强核定位信号(NLS)序列使其从核位置促进转化。只有细胞质定位而非核定位的VirE2能够刺激转化。为了研究VirE2支持转化的方式,我们构建了在诱导型启动子控制下含有一个基因的转基因植物,并在诱导前后进行了RNA测序和蛋白质组分析。一些差异表达的植物基因先前已知可促进转化。其他一些VirE2差异表达基因的敲除突变株表现出改变的转化表型。一些已知对转化重要的蛋白质水平在VirE2诱导后升高,但在其相应mRNA诱导之前或未诱导时就已升高。VirE2诱导后其蛋白质增加的一些其他基因的过表达导致转化敏感性增加。我们得出结论,细胞质定位的VirE2调节植物RNA和蛋白质水平以促进转化。