Tirado-Class Neysha, Hathaway Caitlin, Chung Wendy K, Dungrawala Huzefa
University of South Florida.
Columbia University.
Cold Spring Harb Mol Case Stud. 2022 Jul 21;8(5). doi: 10.1101/mcs.a006212.
Chung-Jansen syndrome (CJS) is a rare, autosomal dominant disorder characterized by developmental delay, intellectual disability/cognitive impairment, behavioral challenges, obesity, and dysmorphic features. CJS is associated with heterozygous variants in PHIP (Pleckstrin-Homology Interacting Protein), a gene that encodes one of several substrate receptors for Cullin4-RING (CRL4) E3 ubiquitin ligase complex. Full length PHIP, also called DCAF14, was recently identified to function as a replication stress response protein. Herein, we report the identification of two PHIP missense variants identified by exome sequencing in unrelated individuals with CJS. The variants p.D488V and p.E963G occur in different functional elements of DCAF14- WD40 repeat domain and pleckstrin homology-binding region (PBR), respectively. Using DNA fiber assays, we reveal that cells expressing either variant exhibit defective replication fork progression in conditions of replication stress. Furthermore, unlike wild type DCAF14, both variants fail to accomplish DNA replication after exposure to genotoxic stress indicating a critical role of DCAF14 in protecting stalled replication forks. Thus, we have identified replication defects associated with CJS variants and predict replication-associated genome instability with CJS syndrome.
钟-扬森综合征(CJS)是一种罕见的常染色体显性疾病,其特征为发育迟缓、智力残疾/认知障碍、行为挑战、肥胖和畸形特征。CJS与PHIP(普列克底物蛋白同源相互作用蛋白)中的杂合变异有关,PHIP是一种基因,编码Cullin4-RING(CRL4)E3泛素连接酶复合体几种底物受体之一。全长PHIP,也称为DCAF14最近被鉴定为一种复制应激反应蛋白。在此我们报告通过外显子组测序在无关的CJS个体中鉴定出两个PHIP错义变异。变异p.D488V和p.E963G分别出现在DCAF14的不同功能元件中——WD40重复结构域和普列克底物蛋白同源结合区域(PBR)。使用DNA纤维测定法我们发现表达任何一种变异的细胞在复制应激条件下均表现出复制叉进展缺陷。此外,与野生型DCAF14不同,两种变异在暴露于遗传毒性应激后均无法完成DNA复制,这表明DCAF14在保护停滞的复制叉中起关键作用。因此,我们已经鉴定出与CJS变异相关的复制缺陷,并预测CJS综合征存在与复制相关的基因组不稳定性。