NUCLEAR MATRIX STRUCTURE AND DNA REPLICATION

核基质结构和 DNA 复制

• 批准号: 2174184
• 负责人: Ronald Berezney
• 金额: $ 29.94万
• 依托单位: STATE UNIVERSITY OF NEW YORK AT BUFFALO
• 依托单位国家: 美国
• 财政年份: 1977
• 资助国家: 美国
• 起止时间: 1977-09-30 至 1999-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2174184
• 关键词: DNA replication; DNA replication origin; affinity chromatography; cell nucleus; chemical binding; chromosomes; density gradient ultracentrifugation; electron microscopy; fluorescence microscopy; gel electrophoresis; human genetic material tag; immunofluorescence technique; in situ hybridization; ion exchange chromatography; laboratory rat; nuclear matrix; nucleic acid sequence; nucleoproteins; protein structure function; scintillation counter; tissue /cell culture

The cell nucleus is the repository for the genetic information of all eucaryotic cells including that of man. Despite considerable progress in defining basic molecular properties of the primary genomic functions of DNA replication, transcription, and RNA splicing and processing, our knowledge of how these processes are organized and regulated within the confines of the cell nucleus is extremely limited. Similarly, although the genetic code behind the DNA (the nucleotide sequence) has long been broken, our understanding of the organization of this DNA into chromatin and higher order structures in the cell nucleus is still in its infancy. Progress in several areas, however, gives one reason to be optimistic about future success. Development of appropriate methods to examine the three dimensional basis of nuclear architecture coupled with new and sensitive detection systems for function, such as fluorescence in situ hybridization (FISH) and fluorescence microscopic detection of DNA replication sites are among the hopes of the future. Our laboratory has been studying the nuclear matrix and its role in DNA replication for over 15 years. Now that basic groundwork implicating the nuclear matrix as a site for the organization of DNA replication has been set, we are now focusing our attention on the 3-D organization of replication in the cell nucleus. By combining these 3-D approaches with the powerful techniques of multi-dimensional computer image analysis and FISH, we are making progress in mapping the DNA replication sites in 3-D. Use of a chromosome 11 set of DNA probes will further enable us to determine the DNA sequence organization of chromosome 11 in 3-D and map the sequences present at individual replication sites. The major directions to be pursued in this project include : (1) Study of the 3-D organization of DNA replication sites in the cell nucleus; (2) Localization of proteins at replication sites in 3-D; (3) 3-D mapping of gene sequences in the cell nucleus for human chromosome 11; (4) Map the replication of genes for human chromosome 11 in 3-D.

细胞核是所有生物遗传信息的储存库 真核细胞，包括人类的细胞。 尽管取得了长足的进步 定义主要基因组功能的基本分子特性 DNA 复制、转录以及 RNA 剪接和加工，我们的 了解这些过程如何在内部组织和监管 细胞核的范围极其有限。 同样，虽然 DNA（核苷酸序列）背后的遗传密码早已被 我们对 DNA 组织成染色质的理解被打破了 细胞核中的高级结构仍处于起步阶段。 然而，几个领域的进展让人有理由感到乐观 关于未来的成功。 制定适当的方法来检查 核结构的三维基础加上新的和 灵敏的功能检测系统，例如原位荧光 DNA 杂交 (FISH) 和荧光显微镜检测 复制位点是未来的希望之一。 我们实验室有 多年来一直在研究核基质及其在 DNA 复制中的作用 15 年。 现在涉及核矩阵的基本基础是 DNA复制的组织位点已经确定，我们现在 将我们的注意力集中在细胞内复制的 3D 组织上 核。 通过将这些 3D 方法与强大的技术相结合 多维计算机图像分析和 FISH，我们正在制作 3-D 绘制 DNA 复制位点的进展。 使用一个 11号染色体DNA探针组将进一步使我们能够确定 11 号染色体的 DNA 序列以 3D 形式组织并绘制序列图 存在于各个复制位点。 拟定的主要方向 本项目的研究内容包括： (1) 3D 组织的研究 细胞核中的DNA复制位点； (2)蛋白质的定位 在 3D 复制位点； (3) 基因序列的 3-D 映射 人类11号染色体的细胞核； (4)绘制基因复制图谱 3D 中的人类 11 号染色体。