Key Laboratory of Systems Biology, State Key Laboratory of Molecular Biology, Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai 200031, China.
Acta Biochim Biophys Sin (Shanghai). 2009 Dec;41(12):991-7. doi: 10.1093/abbs/gmp096.
PES1, the human homolog of zebrafish pescadillo, is a nucleolar protein that is essential for cell proliferation. We report herein that a nucleolar marker protein B23 physically interacts with PES1 and is involved in the nucleolar localization of PES1. In vivo interaction between B23 and PES1 was verified by co-immunoprecipitation of endogenous B23 and PES1 proteins, and they showed cellular co-localizations under both normal and actinomycin D-induced stress conditions. Furthermore, we mapped their interaction domains via in vitro pulldown assays. When B23 was knocked down by RNA interference, there appeared an increased nucleoplasmic distribution of PES1. Our results support a previous hypothesis that B23 might be a nucleolar hub protein for protein targeting to the nucleolus, and shed light on the nucleolar localization mechanism of PES1. The physical interaction between B23 and PES1 implies that they may participate in ribosome biogenesis in a protein complex.
PES1 是人类 pescadillo 的同源物,是一种核仁蛋白,对细胞增殖至关重要。我们在此报告称,核仁标记蛋白 B23 与 PES1 发生物理相互作用,并参与 PES1 的核仁定位。通过共免疫沉淀内源性 B23 和 PES1 蛋白验证了它们在体内的相互作用,并且在正常和放线菌素 D 诱导的应激条件下它们显示出细胞共定位。此外,我们通过体外下拉测定法对它们的相互作用域进行了作图。当 B23 被 RNA 干扰敲低时,PES1 出现核质分布增加。我们的结果支持了 B23 可能是一种核仁枢纽蛋白,用于将蛋白质靶向核仁的假说,并阐明了 PES1 的核仁定位机制。B23 和 PES1 之间的物理相互作用意味着它们可能参与核糖体生物发生的蛋白质复合物。