The large GTPase AtGBPL3 links nuclear envelope formation to lamina-associated gene silencing [ChIP-Seq]

Guanylate Binding Proteins (GBPs) are prominent regulators of immunity unknown to be required for nuclear envelope formation and function. The Arabidopsis GBP orthologue AtGBPL3 was identified as a novel component of the plant lamina, with essential functions in mitotic nuclear envelope reformation. AtGBPL3 is preferentially expressed in mitotically active root tips and accumulates at the nuclear envelope. Furthermore, fluorescent lifetime imaging experiments in the presence or absence of the chromatin-labelling dye Sytox-Orange suggested that GBPL3 might be able to interact with chromatin. Chromatin-immunoprecipitation followed by sequencing (ChIP-Seq) revealed only few GBPL3-associated chromatin domains, which were concentrated particularly within or near centromeric regions. Overall design: Chromatin immunoprecipitation DNA-sequencing (ChIP-seq) for guanylate-binding protein-like 3 (AtGBPL3) from Arabidopsis thaliana

鸟苷酸结合蛋白（Guanylate Binding Proteins，GBPs）是一类重要的免疫调控因子，此前尚未被证实参与核被膜的形成与功能维持。拟南芥的GBP同源蛋白AtGBPL3被鉴定为植物核纤层的新型组分，在有丝分裂核被膜重建过程中发挥不可或缺的功能。AtGBPL3优先在有丝分裂活跃的根尖组织中表达，并定位于核被膜。此外，在添加或不添加染色质标记染料Sytox-Orange的条件下开展的荧光寿命成像（fluorescent lifetime imaging）实验提示，GBPL3可能与染色质存在相互作用。染色质免疫共沉淀测序（Chromatin-immunoprecipitation followed by sequencing，ChIP-Seq）结果显示，仅存在少量与GBPL3结合的染色质结构域，且这些结构域尤其富集于着丝粒区域内部或其邻近位置。实验整体设计：针对来自拟南芥（Arabidopsis thaliana）的鸟苷酸结合蛋白样3（AtGBPL3）开展的染色质免疫共沉淀DNA测序（ChIP-seq）实验。