Imaging files for: ACD15, ACD21, and SLN regulate accumulation and mobility of MBD6 to silence genes and transposable elements

It's well known that DNA methylation is linked to gene silencing but the mechanisms of how proteins that bind the DNA methylation cause gene silencing remains unclear. We demonstrated that the novel MBD5/6 complex contains three chaperone proteins, called ACD15, ACD21, and SLN, which specifically mediate the gene silencing function. ACD15 and ACD21 bridge the interaction of SLN to MBD5 and/or MBD6 while also functioning to drive the accumulation of the MBD5/6 complex at CG methylation sites. We further discovered that SLN also regulates the accumulation of the MBD5/6 complex and regulates the turnover of all protein members once accumulated at meCG sites. 
To demonstrate these results we primarily used fluorescence, confocal microscopy using RFP tagged MBD6, YFP tagged ACD15, and CFP tagg ACD21 and SLN imaging the roots of Arabidopsis thaliana. We expressed these constructs either alone or together in multiple mutant lines including mbd5 mbd6, acd15, acd21, acd15 acd21, sln, and acd15 a...

众所周知，DNA甲基化（DNA methylation）与基因沉默（gene silencing）密切相关，但结合DNA甲基化的蛋白质引发基因沉默的具体机制仍尚不明确。本研究证实，新型MBD5/6复合物（MBD5/6 complex）包含三种伴侣蛋白，分别为ACD15、ACD21与SLN，它们可特异性介导基因沉默功能。ACD15与ACD21一方面介导SLN与MBD5和/或MBD6的相互作用，另一方面还可推动MBD5/6复合物在CG甲基化位点的富集。我们进一步发现，SLN不仅可调控MBD5/6复合物的富集过程，还可调控该复合物在甲基化CG（meCG）位点积累后所有蛋白组分的周转过程。
为验证上述研究结果，本研究主要采用荧光共聚焦显微镜技术，对红色荧光蛋白（Red Fluorescent Protein, RFP）标记的MBD6、黄色荧光蛋白（Yellow Fluorescent Protein, YFP）标记的ACD15以及青色荧光蛋白（Cyan Fluorescent Protein, CFP）标记的ACD21与SLN在拟南芥（Arabidopsis thaliana）根部的表达情况进行成像分析。我们将这些构建体分别或共同在多种突变体株系中表达，包括mbd5 mbd6、acd15、acd21、acd15 acd21、sln以及acd15 a...