National Laboratory of Biomacromolecules, CAS Center for Excellence in Biomacromolecules, Institute of Biophysics, Chinese Academy of Sciences, Beijing, 100101, P. R. China.
College of Life Sciences, University of Chinese Academy of Sciences, Beijing, 100049, China.
Sci Rep. 2017 Nov 15;7(1):15638. doi: 10.1038/s41598-017-15715-9.
CDC73/Parafibromin is a critical component of the Paf1 complex (PAF1C), which is involved in transcriptional elongation and histone modifications. Mutations of the human CDC73/HRPT2 gene are associated with hyperparathyroidism-jaw tumor (HPT-JT) syndrome, an autosomal dominant disorder. CDC73/parafibromin was initially recognized as a tumor suppressor by inhibiting cell proliferation via repression of cyclin D1 and c-myc genes. In recent years, it has also shown oncogenic features by activating the canonical Wnt/β-catenin signal pathway. Here, through limited proteolysis analysis, we demonstrate that the evolutionarily conserved human CDC73 N-terminal 111 residues form a globularly folded domain (hCDC73-NTD). We have determined a crystal structure of hCDC73-NTD at 1.02 Å resolution, which reveals a novel protein fold. CDC73-NTD contains an extended hydrophobic groove on its surface that may be important for its function. Most pathogenic CDC73 missense mutations associated with the HPT-JT syndrome are located in the region encoding CDC73-NTD. Our crystal and biochemical data indicate that most CDC73 missense mutations disrupt the folding of the hydrophobic core of hCDC73-NTD, while others such as the K34Q mutant reduce its thermostability. Overall, our results provide a solid structural basis for understanding the structure and function of CDC73 and its association with the HPT-JT syndrome and other diseases.
CDC73/Parafibromin 是 Paf1 复合物(PAF1C)的关键组成部分,该复合物参与转录延伸和组蛋白修饰。人类 CDC73/HRPT2 基因突变与甲状旁腺功能亢进-颌骨肿瘤(HPT-JT)综合征有关,这是一种常染色体显性遗传病。CDC73/parafibromin 最初被认为是一种肿瘤抑制因子,通过抑制细胞增殖来抑制 cyclin D1 和 c-myc 基因的表达。近年来,它还通过激活经典的 Wnt/β-catenin 信号通路表现出致癌特征。在这里,通过有限的蛋白水解分析,我们证明了进化上保守的人类 CDC73 N 端 111 个残基形成一个球状折叠结构域(hCDC73-NTD)。我们已经确定了 hCDC73-NTD 的晶体结构,分辨率为 1.02 Å,揭示了一种新的蛋白质折叠。CDC73-NTD 表面有一个伸展的疏水性凹槽,这可能对其功能很重要。与 HPT-JT 综合征相关的大多数致病性 CDC73 错义突变位于编码 CDC73-NTD 的区域。我们的晶体和生化数据表明,大多数 CDC73 错义突变破坏了 hCDC73-NTD 疏水性核心的折叠,而其他突变,如 K34Q 突变,则降低了其热稳定性。总的来说,我们的结果为理解 CDC73 的结构和功能及其与 HPT-JT 综合征和其他疾病的关联提供了坚实的结构基础。