Control of the mid-blastula transition in Drosophila

果蝇中囊胚转变的控制

• 批准号: 7152718
• 负责人: Mark Lawrence McCleland
• 金额: $ 4.88万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-07-01 至 2008-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7152718
• 关键词: DNA replication; Drosophilidae; cell cycle; chromatin; cyclin dependent kinase; cyclins; cytogenetics; developmental genetics; embryo /fetus; embryogenic cleavage; genetic regulation; histones; in situ hybridization; invertebrate embryology; mother /embryo /fetus nutrition; postdoctoral investigator; posttranslational modifications

DESCRIPTION (provided by applicant): In the 14th division of the Drosophila embryo, the rapid maternally driven cycles end and zygotic regulation begins. This transition, referred to as the mid-blastula transition (MBT), coordinates a multitude of cellular events to ensure developmental fidelity. This proposal seeks to characterize how titration of cyclin:Cdk activity and changes in chromatin modifications influence the MBT. By genetically altering cyclin:cdk activity, we will monitor: cell cycle timing by real time flourescence microscopy, replication dynamics by injecting labeled nucleotides, and the transcription of key genes by in situ hybridization. Continued characterization of the "histone code" will define changes in chromatin modifications at the MBT and injecting deacetlyase inhibitors into staged embryos will directly test how chromatin state influences the MBT and replication. Finally, the relationship between cyclin:Cdk activity and histone modifications will be examined using mutants of cyclin:Cdk regulation, non-degradable cyclins, and Cdk inhibitors. Understanding the MBT and cell cycle transitions is important not only in discerning basic development, but in elucidating how defects in this process lead to abnormalities, including cancer and birth defects.

描述（由申请人提供）：在果蝇胚胎的第14次分裂中，快速的母系驱动周期结束，合子调节开始。这种转变，被称为中期囊胚转变（MBT），协调大量的细胞事件，以确保发育的保真度。该建议旨在表征细胞周期蛋白：Cdk活性的滴定和染色质修饰的变化如何影响MBT。通过基因改变细胞周期蛋白：cdk活性，我们将监测：细胞周期的时间通过真实的荧光显微镜，复制动力学通过注射标记核苷酸，和转录的关键基因通过原位杂交。继续表征“组蛋白密码”将定义MBT处染色质修饰的变化，将脱乙酰酶抑制剂注射到分期胚胎中将直接测试染色质状态如何影响MBT和复制。最后，细胞周期蛋白：Cdk活性和组蛋白修饰之间的关系将使用细胞周期蛋白：Cdk调节，不可降解的细胞周期蛋白，和Cdk抑制剂的突变体进行检查。了解MBT和细胞周期转换不仅对识别基本发育很重要，而且对阐明这一过程中的缺陷如何导致异常，包括癌症和出生缺陷也很重要。