Institute of Human Genetics, Charité - Universitätsmedizin Berlin, Berlin, Germany.
Cell Cycle. 2011 Sep 1;10(17):2967-77. doi: 10.4161/cc.10.17.16871.
Primary autosomal recessive microcephaly (MCPH) is a congenital disorder characterized by a pronounced reduction of brain size and mental retardation. We present here a consanguineous Turkish family clinically diagnosed with MCPH and without linkage to any of the known loci (MCPH1-MCPH7). Autozygosity mapping identified a homozygous region of 15.8 Mb on chromosome 10q11.23-21.3, most likely representing a new locus for MCPH. Although we were unable to identify the underlying genetic defect after extensive molecular screening, we could delineate a possible molecular function in chromosome segregation by the characterization of mitosis in the patients' cells. Analyses of chromosome nondisjunction in T-lymphocytes and fibroblasts revealed a significantly elevated rate of nondisjunction in the patients' cells as compared to controls. Mitotic progression was further explored by immunofluorescence analyses of several chromosome and spindle associated proteins. We detected a remarkable alteration in the anaphase distribution of Aurora B and INCENP, which are key regulators of chromosome segregation. In particular, a fraction of both proteins remained abnormally loaded on chromosomes during anaphase in MCPH patients' cells while in cells of normal control subjects both proteins are completely transferred to the spindle midzone. We did not observe any other alterations regarding cell cycle progression, chromosome structure, or response to DNA damage. Our observations point towards a molecular role of the underlying gene product in the regulation of anaphase/telophase progression possibly through interaction with chromosomal passenger proteins. In addition, our findings represent further evidence for the proposed role of MCPH genes in the regulation of mitotic progression.
原发性常染色体隐性小头畸形(MCPH)是一种先天性疾病,其特征为大脑体积明显缩小和智力迟钝。我们在此介绍一个土耳其的近亲家族,他们被临床诊断为 MCPH,但与已知的任何基因座(MCPH1-MCPH7)均无连锁关系。单体型分析确定了 10q11.23-21.3 染色体上的一个 15.8Mb 的纯合区域,该区域很可能代表 MCPH 的新基因座。尽管我们在进行广泛的分子筛选后仍未能确定潜在的遗传缺陷,但通过对患者细胞有丝分裂的特征分析,我们可以描绘出染色体分离的可能分子功能。对 T 淋巴细胞和成纤维细胞的染色体非整倍体分析显示,与对照组相比,患者细胞的非整倍体率显著升高。通过对几种染色体和纺锤体相关蛋白的免疫荧光分析进一步研究了有丝分裂的进展。我们发现 Aurora B 和 INCENP 的后期分布发生了显著改变,这两种蛋白都是染色体分离的关键调节因子。特别是,在 MCPH 患者的细胞中,这两种蛋白的一部分在后期仍异常加载在染色体上,而在正常对照细胞中,这两种蛋白均完全转移到纺锤体中部。我们没有观察到与细胞周期进程、染色体结构或对 DNA 损伤的反应有关的任何其他改变。我们的观察结果表明,潜在基因产物在调节后期/末期进展中具有分子作用,可能通过与染色体乘客蛋白相互作用。此外,我们的发现进一步证明了 MCPH 基因在调节有丝分裂进程中的作用。