METAMORPHOSIS OF CHROMATIN

染色质变态

• 批准号: 3303648
• 负责人: JUDITH WILLIS
• 金额: $ 12.02万
• 依托单位: UNIVERSITY OF GEORGIA
• 依托单位国家: 美国
• 财政年份: 1990
• 资助国家: 美国
• 起止时间: 1990-07-01 至 1995-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3303648
• 关键词: DNA binding protein; DNA footprinting; Lepidoptera; binding proteins; chromatin; complementary DNA; deoxyribonuclease I; developmental genetics; endonuclease; gel mobility shift assay; gene expression; genetic manipulation; genetic regulation; genetic regulatory element; genome; homeobox genes; hormone receptor; hormone regulation /control mechanism; invertebrate cuticle; laboratory mouse; metamorphosis; molecular genetics; nucleic acid hybridization; nucleic acid probes; nucleic acid sequence; polymerase chain reaction; protein biosynthesis; protein sequence; DNA binding protein; DNA footprinting; Lepidoptera; binding proteins; chromatin; complementary DNA; deoxyribonuclease I; developmental genetics; endonuclease; gel mobility shift assay; gene expression; genetic manipulation; genetic regulation; genetic regulatory element; genome; homeobox genes; hormone receptor; hormone regulation /control mechanism; invertebrate cuticle; laboratory mouse; metamorphosis; molecular genetics; nucleic acid hybridization; nucleic acid probes; nucleic acid sequence; polymerase chain reaction; protein biosynthesis; protein sequence

Metamorphosis in moths involves a continuous population of cells changing their synthetic activities to build new types of cuticle under the direct influence of two hormones, ecdysteroids and juvenoids. The changes that occur and our ability to manipulate them in vivo and in vitro presents a useful model for analyzing developmental events in our parasites and ourselves. We propose to learn how metamorphosis is regulated at the nomic level. To this end we shall isolate and characterize genes and transcripts for four cuticular proteins selected for their provocative spatial and temporal distributions. We plan to analyze chromatin to identify the regulatory domains that encompass these genes. Putative cis-acting regulatory regions will be located by using DNase I to map hypersensitive sites and by using dimethylsulfate (DMS) for "in vivo footprinting". Putative regulatory regions identified by these techniques will be used in gel retardation assays where the presence of regulatory proteins can be recognized since they reduce the mobility of DNA to which they are specifically bound. We shall test nuclear extracts from different anatomical regions and metamorphic stages as well as known trans-acting regulatory factors to sort out the nature (spatial, temporal, hormonal) of the information impinging on the cis-acting regulatory sequences of a single gene during the course of metamorphosis. We hope by these analyses to learn if hormones control metamorphosis by acting directly ont he genes that code for structural proteins. The moth epidermis presents an extremely favorable model for analyzing the fundamental issue of how chromatin structure is altered as a continuous population of cells plays out a development program that involves changes in the state of determination and activity for individual genes.

飞蛾中的变形涉及连续的细胞群体变化 他们在直接下建立新型角质层的合成活动 两种激素，雌激素和青少年的影响。 变化 发生以及我们在体内和体外呈现A的能力 用于分析我们寄生虫的发展事件的有用模型 我们自己。 我们建议学习在杂志上如何调节变形 等级。 为此，我们将隔离并表征基因和转录本 对于四个用于挑衅性空间的表皮蛋白和 时间分布。 我们计划分析染色质以识别 包含这些基因的调节域。 推定的顺式作用 调节区域将通过使用DNase I绘制超敏敏感 位置和通过使用二甲基硫酸盐（DMS）进行“体内足迹”。 这些技术确定的推定监管区域将用于 凝胶延迟测定可能是调节蛋白的存在 由于它们降低了DNA的迁移率，因此被认可 特别绑定。 我们将测试来自不同的核提取物 解剖区域和变质阶段以及已知的反式作用 调节因素，以分析性质（空间，时间，荷尔蒙） 撞击顺式作用调节序列的信息 在变形过程中单基因。我们希望通过这些分析 了解激素是否通过直接作用于基因来控制变形 结构蛋白的代码。 飞蛾表皮提出了 非常有利的模型，用于分析如何 随着细胞的连续群体的播放，染色质结构被改变 制定涉及变化状态的开发计划 单个基因的确定和活性。