Differential DNA Replication in Drosophila Development

果蝇发育中的差异DNA复制

• 批准号: 8071619
• 负责人: Terry L. ORR-WEAVER
• 金额: $ 39.18万
• 依托单位: WHITEHEAD INSTITUTE FOR BIOMEDICAL RES
• 依托单位国家: 美国
• 财政年份: 1999
• 资助国家: 美国
• 起止时间: 1999-05-01 至 2012-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8071619
• 关键词: Affect; Animals; Binding; Binding Proteins; Binding Sites; Cell Cycle; Cell Differentiation process; Cells; Cellular biology; Collection; DNA; DNA biosynthesis; DNA replication origin; Development; Drosophila genus; E2F transcription factors; Ensure; Factor Analysis; Foundations; Funding; Gene Dosage; Genes; Genetic Transcription; Genomics; Goals; Histone Acetylation; Human; Metastatic to; Methodology; Mitosis; Modeling; Monitor; Movement; Ovarian Follicle; Ploidies; Polyploid Cells; Polyploidy; Process; Protein Binding; Regulation; Replication Origin; Repression; Research; Role; Salivary Glands; Staging; Structure; Tissues; genetic regulatory protein; mutant; protein function; tumor progression; Affect; Animals; Binding; Binding Proteins; Binding Sites; Cell Cycle; Cell Differentiation process; Cells; Cellular biology; Collection; DNA; DNA biosynthesis; DNA replication origin; Development; Drosophila genus; E2F transcription factors; Ensure; Factor Analysis; Foundations; Funding; Gene Dosage; Genes; Genetic Transcription; Genomics; Goals; Histone Acetylation; Human; Metastatic to; Methodology; Mitosis; Modeling; Monitor; Movement; Ovarian Follicle; Ploidies; Polyploid Cells; Polyploidy; Process; Protein Binding; Regulation; Replication Origin; Repression; Research; Role; Salivary Glands; Staging; Structure; Tissues; genetic regulatory protein; mutant; protein function; tumor progression

DESCRIPTION (provided by applicant): Control of DNA replication is critical to ensure accurate copy number of genes, because both loss of gene copies and increased copy number by amplification are associated with the onset of cancer and progression to metastatic states. Elucidation of regulatory mechanisms for metazoan DNA replication has been hampered by difficulty in defining specific origins of replication and monitoring their activation. Most animals, including humans, contain polyploid tissues in which the DNA content of the cells is increased by a modified cell cycle, the endo cycle, lacking mitosis. In addition to the overall increase in genomic DNA, in many polyploid cells differential DNA replication occurs in which specific genomic intervals are not replicated or in some cases are over-replicated and amplified. These instances of differential DNA replication provide superb models for elucidating the structure and regulation of metazoan DNA replication origins. Using genomic methodologies, amplified genomic regions were identified in the Drosophila ovarian follicle cells and single copy, euchromatic underreplicated regions were found in the larval salivary gland. The replication origins within the amplified regions are subject to developmental control and permit analysis of the factors responsible for origin activation and repression, as well as discovery of mechanisms by which metazoan DNA replication is initiated. The aims of this research are to exploit these Drosophila replication models to define the mechanisms that activate initiation at the origins during follicle cell differentiation, to determine how replication origins are inactivated and to use mutants to identify new regulatory proteins, and to analyze mechanisms controlling replication fork progression. The identification of defined replication origins whose duplication can be quantified, the ability to detect replication proteins bound at these origins and moving with the replication forks, combined with a collection of mutants affecting these processes permits these experimental goals to be achieved.

描述（由申请人提供）：对DNA复制的控制对于确保基因的准确拷贝数至关重要，因为基因副本的丢失和通过扩增增加的拷贝数增加与癌症的发作和向转移状态的发展有关。难以确定复制的特定起源和监测其激活的困难，阐明了后生DNA复制的调节机制。包括人类在内的大多数动物都包含多倍体组织，其中细胞的DNA含量通过修饰的细胞周期，内部周期增加，缺乏有丝分裂。除了基因组DNA的总体增加外，在许多多倍体细胞中，发生了差异DNA复制，其中未重复特定的基因组间隔，或者在某些情况下被过度复制和放大。这些差异DNA复制的实例提供了出色的模型，以阐明后生DNA复制起源的结构和调节。使用基因组方法，在果蝇卵巢卵泡细胞中鉴定出扩增的基因组区域和单个拷贝，在幼虫唾液腺中发现了构型均匀复制的区域。放大区域内的复制起源受发展控制和允许分析负责起源激活和抑制的因素，以及发现启动后生DNA复制的机制。这项研究的目的是利用这些果蝇复制模型来定义在卵泡细胞分化过程中激活起源的机制，以确定复制起源如何灭活，并使用突变体识别新的调节蛋白，并分析控制复制机制的机制。可以量化重复的定义复制起源的识别，可检测的能力 复制蛋白以这些起源结合并与复制叉一起移动，结合一系列影响这些过程的突变体可以实现这些实验目标。