Sukumaran Salil K, Blau-Wasser Rosemarie, Rohlfs Meino, Gallinger Christoph, Schleicher Michael, Noegel Angelika A
a Institute of Biochemistry I; Medical Faculty University of Cologne ; Köln , Germany.
Cell Cycle. 2015;14(7):1024-35. doi: 10.1080/15384101.2015.1007015.
CEP161 is a novel component of the Dictyostelium discoideum centrosome which was identified as binding partner of the pericentriolar component CP250. Here we show that the amino acids 1-763 of the 1381 amino acids CEP161 are sufficient for CP250 binding, centrosomal targeting and centrosome association. Analysis of AX2 cells over-expressing truncated and full length CEP161 proteins revealed defects in growth and development. By immunoprecipitation experiments we identified the Hippo related kinase SvkA (Hrk-svk) as binding partner for CEP161. Both proteins colocalize at the centrosome. In in vitro kinase assays the N-terminal domain of CEP161 (residues 1-763) inhibited the kinase activity of Hrk-svk. A comparison of D. discoideum Hippo kinase mutants with mutants overexpressing CEP161 polypeptides revealed similar defects. We propose that the centrosomal component CEP161 is a novel player in the Hippo signaling pathway and affects various cellular properties through this interaction.
CEP161是盘基网柄菌中心体的一个新组分,它被鉴定为中心粒外周组分CP250的结合伴侣。在此我们表明,1381个氨基酸的CEP161中的1至763位氨基酸足以实现CP250结合、中心体靶向定位和中心体缔合。对过表达截短型和全长CEP161蛋白的AX2细胞进行分析,发现其生长和发育存在缺陷。通过免疫沉淀实验,我们鉴定出与Hippo相关的激酶SvkA(Hrk-svk)为CEP161的结合伴侣。这两种蛋白在中心体处共定位。在体外激酶分析中,CEP161的N端结构域(第1至763位氨基酸残基)抑制了Hrk-svk的激酶活性。将盘基网柄菌Hippo激酶突变体与过表达CEP161多肽的突变体进行比较,发现了类似的缺陷。我们提出,中心体组分CEP161是Hippo信号通路中的一个新成员,并通过这种相互作用影响多种细胞特性。