MAINTAINING DNA REPLICATION FORK STABILITY: ROLE OF THE FANCONI ANEMIA PATHWAY

维持 DNA 复制叉稳定性：范可尼贫血途径的作用

• 批准号: 7960146
• 负责人: Niall George Howlett
• 金额: $ 19.96万
• 依托单位: UNIVERSITY OF RHODE ISLAND
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-05-04 至 2010-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7960146
• 关键词: Aphidicolin; Behavioral Research; Cells; Chromosome Breakage; Chromosome Fragile Sites; Cisplatin; Clinical; Computer Retrieval of Information on Scientific Projects Database; DNA; DNA Crosslinking Agent; DNA Integration; DNA Polymerase Inhibitor; DNA biosynthesis; DNA replication fork; DNA-Directed DNA Polymerase; Disease; FANCD2 protein; FHIT gene; Fanconi' s Anemia; Frequencies; Funding; Genomics; Grant; Human Papillomavirus; Institution; Lead; Maintenance; Malignant Neoplasms; Maps; Mitomycins; Mono-S; Mutation; Neoplastic Cell Transformation; Pancytopenia; Pathway interactions; Patients; Physiological; Play; Predisposition; Prevention; Research; Research Personnel; Resources; Role; Site; Source; Ubiquitination; United States National Institutes of Health; in vivo; insight; response; tumor

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Fanconi anemia (FA) is a rare recessive disorder characterized by congenital anomalies, bone marrow failure, and pronounced cancer susceptibility. FA patient cells are hypersensitive to DNA crosslinking agents such as mitomycin C and cisplatin. The endogenous function of the FA pathway, however, remains unknown. We have recently demonstrated that the FA pathway is functionally required for the cellular response to disruption of DNA replication. The FA pathway is strongly activated via the mono-ubiquitination of the FANCD2 protein following treatment with the DNA polymerase inhibitor aphidicolin (APH). Furthermore, abrogation of the FA pathway leads to increased chromosome breakage, including breakage at the common chromosomal fragile sites FRA3B and FRA16D, following treatment with APH. We hypothesize that the FA pathway plays an integral role in the maintenance of DNA replication fork stability, a function critical for the prevention of neoplastic transformation. Two aims are proposed to gain further insight into this function. First, we will characterize the interaction between the FANCD2 protein and the major DNA polymerase processivity factor PCNA. Specifically, we will examine the functional consequences of mutation of a recently identified FANCD2 PCNA-interaction motif. Second, we will determine if the increased APH-induced chromosome breakage in FA cells leads to increased integration of foreign DNA. Finally, we will attempt to establish a clinical corollary for a role of the FA pathway in the in the maintenance of DNA replication fork stability: We will determine if the increased common chromosomal fragile site breakage of FA cells is associated with an increased frequency of human papillomavirus (HPV) DNA integration at these loci, by mapping the genomic sites of HPV integration in FA patient tumors. Our proposed studies will lead to new findings regarding the in vivo physiological role of the FA pathway, and provide insight into the increased cancer susceptibility of FA patients.

该副本是利用众多研究子项目之一 由NIH/NCRR资助的中心赠款提供的资源。子弹和 调查员（PI）可能已经从其他NIH来源获得了主要资金， 因此可以在其他清晰的条目中代表。列出的机构是 对于中心，这不一定是调查员的机构。 Fanconi贫血（FA）是一种罕见的隐性疾病，其特征是先天性异常，骨髓衰竭和明显的癌症敏感性。 FA患者细胞对DNA交联剂（例如丝裂霉素C和顺铂）过敏。但是，FA途径的内源性功能仍然未知。我们最近证明，FA途径在功能上是对DNA复制破坏的细胞反应所必需的。通过用DNA聚合酶抑制剂蚜虫（APH）处理，通过fancd2蛋白的单素化来强烈激活FA途径。此外，废除FA途径会导致染色体破裂增加，包括在用APH处理后，在常见的染色体脆弱位点FRA3B和FRA16D上断裂。我们假设FA途径在维持DNA复制叉稳定性中起着不可或缺的作用，这对于预防肿瘤转化至关重要。提出了两个目标，以进一步了解此功能。首先，我们将表征FANCD2蛋白与主要DNA聚合酶加工性因子PCNA之间的相互作用。具体而言，我们将检查最近鉴定出的FACD2 PCNA相互作用基序突变的功能后果。其次，我们将确定FA细胞中APH诱导的染色体断裂是否增加导致外源DNA的整合增加。最后，我们将尝试在维持DNA复制叉稳定性中建立临床推论，以实现FA途径的作用：我们将确定FA细胞的常见染色体脆弱的位点破裂是否与人类乳头状瘤病毒（HPV）DNA在这些位置的DNA集成的频率增加有关，该位置通过这些位点绘制基因组的STERATION HPV fa fa fa。我们提出的研究将导致有关FA途径的体内生理作用的新发现，并深入了解FA患者的癌症敏感性增加。