SMARCAL1 function in replication stress and disease

描述(申请人提供)：癌细胞在遗传上不稳定，通常由于复制压力而含有高水平的DNA损伤。这种与正常细胞的差异为使用针对DNA损伤反应(DDR)和其他基因组维持途径的合成致死方法进行治疗干预提供了机会。识别关键的DDR蛋白，确定它们在基因组维持途径中的功能，并确定它们对生存至关重要的背景是至关重要的。拟议的研究将检查SMARCAL1的基因组维持活动，SMARCAL1是一种最近发现的DDR蛋白。SMARCAL1是一个SNF2家族的ATPase，通过与分叉的DNA结构结合而激活。SMARCAL1具有不寻常的退火解旋酶活性，并在停滞的复制叉处发挥作用。我们的初步数据支持这样的假设，即SMARCAL1与其他DDR蛋白一起在DNA复制压力下维持基因组完整性，从而预防疾病。我们将通过完成三个特定的目标来验证这一假设：(1)表征SMARCAL1与特定DDR蛋白的相互作用并确定这些相互作用的功能；(2)评估SMARCAL1在体内响应DNA复制压力和损伤的功能；以及(3)表征SMARCAL1在响应癌基因诱导的复制压力中的作用。提出了创新的生化和遗传方法，包括一种独特的SMARCAL1小鼠模型-功能丧失。该项目的完成将确定SMARCAL1与DDR蛋白相互作用的功能后果，以及SMARCAL1在体内响应药物和癌基因诱导的复制压力的功能。重要的是，该项目结合了两名调查人员的互补专业知识，通过合作，将提供对复制压力和DDR的关键洞察，并将确定新的 在癌症治疗中开拓的治疗途径。

DESCRIPTION (provided by applicant): Cancer cells are genetically unstable and often contain high levels of DNA damage due to replication stress. This difference with normal cells presents an opportunity for therapeutic intervention using synthetic lethal approaches targeting the DNA damage response (DDR) and other genome maintenance pathways. Identification of key DDR proteins, characterizing their function in genome maintenance pathways, and defining the context where they become essential for viability is critical. The proposed studies will examine the genome maintenance activities of SMARCAL1, a recently discovered DDR protein. SMARCAL1 is an SNF2 family of ATPases that is activated by binding to forked DNA structures. SMARCAL1 has an unusual annealing helicase activity and functions at stalled replication forks. Our preliminary data support the hypothesis that SMARCAL1 functions with other DDR proteins to maintain genome integrity during DNA replication stress and thereby prevent disease. We will test this hypothesis by completing three specific aims: (1) characterize the interactions of SMARCAL1 with specific DDR proteins and determine the function of these interactions; (2) evaluate the in vivo function of SMARCAL1 in the response to DNA replication stress and damage; and (3) characterize the role of SMARCAL1 in responding to oncogene-induced replication stress. Innovative biochemical and genetic approaches including a unique mouse model of SMARCAL1-loss of function are proposed. Completion of this project will define the functional consequences of SMARCAL1 interaction with DDR proteins, and the in vivo function of SMARCAL1 in responding to both drug and oncogene-induced replication stress. Importantly, the project combines the complementary expertise of two investigators, who by working together, will provide critical insight into replication stress and the DDR and will identify novel therapeutic avenues to exploit in the treatment of cancer.