Sukumaran Salil K, Stumpf Maria, Salamon Sarah, Ahmad Ilyas, Bhattacharya Kurchi, Fischer Sarah, Müller Rolf, Altmüller Janine, Budde Birgit, Thiele Holger, Tariq Muhammad, Malik Naveed Altaf, Nürnberg Peter, Baig Shahid Mahmood, Hussain Muhammad Sajid, Noegel Angelika A
Institute of Biochemistry I, Medical Faculty, University of Cologne, Joseph-Stelzmann-Str. 52, 50931, Köln, Germany.
Center for Molecular Medicine Cologne (CMMC), University of Cologne, 50931, Köln, Germany.
Mol Genet Genomics. 2017 Apr;292(2):365-383. doi: 10.1007/s00438-016-1277-x. Epub 2016 Dec 21.
Autosomal recessive primary microcephaly (MCPH) is characterized by a substantial reduction in brain size but with normal architecture. It is often linked to mutations in genes coding for centrosomal proteins; however, their role in brain size regulation is not completely understood. By combining homozygosity mapping and whole-exome sequencing in an MCPH family from Pakistan, we identified a novel mutation (XM_011518861.1; c.4114C > T) in CDK5RAP2, the gene associated with primary microcephaly-3 (MCPH3), leading to a premature stop codon (p.Arg1372*). CDK5RAP2 is a component of the pericentriolar material important for the microtubule-organizing function of the centrosome. Patient-derived primary fibroblasts had strongly decreased CDK5RAP2 amounts, showed centrosomal and nuclear abnormalities and exhibited changes in cell size and migration. We further identified an interaction of CDK5RAP2 with the Hippo pathway components MST1 kinase and the transcriptional regulator TAZ. This finding potentially provides a mechanism through which the Hippo pathway with its roles in the regulation of centrosome number is linked to the centrosome. In the patient fibroblasts, we observed higher levels of TAZ and YAP. However, common target genes of the Hippo pathway were downregulated as compared to the control with the exception of BIRC5 (Survivin), which was significantly upregulated. We propose that the centrosomal deficiencies and the altered cellular properties in the patient fibroblasts can also result from the observed changes in the Hippo pathway components which could thus be relevant for MCPH and play a role in brain size regulation and development.
常染色体隐性原发性小头畸形(MCPH)的特征是脑容量显著减小,但结构正常。它通常与编码中心体蛋白的基因突变有关;然而,它们在脑容量调节中的作用尚未完全明确。通过对来自巴基斯坦的一个MCPH家族进行纯合性定位和全外显子组测序,我们在与原发性小头畸形3型(MCPH3)相关的基因CDK5RAP2中发现了一个新的突变(XM_011518861.1;c.4114C>T),该突变导致提前出现终止密码子(p.Arg1372*)。CDK5RAP2是中心粒周围物质的一个组成部分,对中心体的微管组织功能很重要。患者来源的原代成纤维细胞中CDK5RAP2的量大幅减少,表现出中心体和细胞核异常,并出现细胞大小和迁移的变化。我们进一步确定了CDK5RAP2与Hippo信号通路成分MST1激酶和转录调节因子TAZ之间的相互作用。这一发现可能提供了一种机制,通过该机制,在中心体数量调节中起作用的Hippo信号通路与中心体联系起来。在患者的成纤维细胞中,我们观察到TAZ和YAP的水平较高。然而,与对照组相比,Hippo信号通路的常见靶基因除了BIRC5(存活素)显著上调外均下调。我们认为,患者成纤维细胞中的中心体缺陷和细胞特性改变也可能是由于观察到的Hippo信号通路成分变化所致,因此这些变化可能与MCPH相关,并在脑容量调节和发育中发挥作用。