CREB BINDING PROTEIN AND GENE REGULATION

CREB ​​结合蛋白和基因调控

• 批准号: 2843527
• 负责人: WILLIAM H LUDLAM
• 金额: $ 12.66万
• 依托单位: OREGON HEALTH & SCIENCE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1999
• 资助国家: 美国
• 起止时间: 1999-06-01 至 2004-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2843527
• 关键词: Adenoviridae; Drosophilidae; acetyl coA; acyltransferase; binding sites; biological signal transduction; cAMP response element binding protein; chromatin; enzyme activity; gene deletion mutation; genetic regulation; histones; point mutation; protein sequence; virus protein

The cAMP mediated second messenger pathway is a well established model of gene regulation. Within a given cell, many transcription factors are known to interact with and work through the protein kinase A/CREB pathway making integration of multiple signals through common regulatory factors an exceedingly complex phenomenon. The mechanism by which CBP (a CREB co-activator) influences gene regulation is not fully understood, but it is likely that this factor contributes to transcriptional signal integration. There is increasing evidence that this activity may be, in part, due to histone acetylation (HAT) of chromatin that exposes DNA for transcription. The main thrust of this project is to characterize the intrinsic HAT activity of CBP/p300. Four Specific Aims are proposed: (1) CBP/p300 proteins mutated in the putative intrinsic HAT domain (between the bromodomain and the glutamine-rich region of CBP/p300) will be developed to determine and test whether they retain their intrinsic HAT activity. This will also be helpful in mapping the HAT domain. (2) To test whether the intrinsic HAT activity of CBP/p300 for nucleosomes and polynucleosomes is blocked by E1A, since the mechanism by which adenovirus 12S E1A blocks the CBP/p300 co-activator is unknown. (3) To determine whether mutants lose their ability to activate reconstituted chromatin (DNA bound to histones), yet maintain their ability to activate transcription in DNA not bound to histones. (4) To study CBP HAT mutants in developing flies. This lab has previously shown that dCBP rescues CBP-null flies. The inability of CBP HAT mutants to rescue CBP-null flies would be strong evidence that intrinsic HAT activity of CBP (and p300) is critical for in vivo function, and that histone acetylation plays a central role in their gene regulatory function. Gaining insight to the mechanisms of gene activation and silencing has far reaching relevance in the treatment of many diseases including cancers and endocrinopathies. Through a mentored career development plan, the Principal Investigator will gain an enhanced capacity to be productive in the field of molecular biology with a focus on gene regulation. This will be accomplished by conducting the above described research, attending and presenting at local and national seminars and meetings, and participating in courses teaching molecular biology techniques. The proposed mentoring laboratory and institution are rich in intellectual and physical facility resources that provide an excellent environment for the Principal Investigator to transition to independence in the design and implementation of research in the field of gene regulation.

cAMP介导的第二信使途径是基因调控的良好模型。 在给定的细胞中，已知许多转录因子与蛋白激酶A/CREB途径相互作用并起作用，从而使多个信号通过常见的调节因子整合成为非常复杂的现象。 CBP（CREB共激活因子）影响基因调节的机制尚不完全了解，但是该因素可能有助于转录信号整合。 越来越多的证据表明，这种活性可能部分是由于染色质的组蛋白乙酰化（HAT）导致DNA进行转录。 该项目的主要目的是表征CBP/p300的固有HAT活动。 提出了四个具体目的：（1）将开发和测试他们是否保留其内在的HAT活性，以确定和测试它们是否保留其内在的HAT活性。 这也将有助于映射帽子域。 （2）测试CBP/p300对核小体和多核体的固有HAT活性是否被E1A阻断，因为尚不清楚腺病毒12S E1A阻止CBP/P300共激活剂的机制是未知的。 （3）确定突变体是否失去了激活重构染色质（与组蛋白结合的DNA）的能力，但保持其激活与组蛋白不结合的DNA中转录的能力。 （4）在发育中研究CBP帽子突变体。该实验室先前已经表明DCBP营救了CBP-Null Fly。 CBP HAT突变体无法营救CBP-NULL FLIE，这是有力的证据表明，CBP（和p300）的内在HAT活性对于体内功能至关重要，并且组蛋白乙酰化在其基因调节功能中起着核心作用。 洞悉基因激活和沉默的机制与包括癌症和内分泌病在内的许多疾病的治疗相关。 通过指导的职业发展计划，首席研究人员将获得增强的能力，以在分子生物学领域提高生产力，重点是基因调节。 这将通过上述研究，参加并在本地和国家研讨会和会议上进行，并参加教学分子生物学技术的课程来实现。 拟议的指导实验室和机构拥有丰富的智力和物理设施资源，为首席研究人员提供了一个极好的环境，使其在基因监管领域的研究和实施方面过渡到独立性。