MECHANISMS OF TRANSCRIPTIONAL COORDINATION AMONG PHOSPHORYLASE KINASE GENES

磷酸化酶激酶基因之间的转录协调机制

• 批准号: 7381776
• 负责人: Nancy Ayers Rice
• 金额: $ 12.19万
• 依托单位: UNIVERSITY OF LOUISVILLE
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-05-01 至 2007-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7381776
• 关键词: MECHANISMS; TRANSCRIPTIONAL; COORDINATION; AMONG; PHOSPHORYLASE

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Phosphorylase kinase (PhK) is a key regulatory enzyme involved in the glycogenolytic pathway, catalyzing the activation, by phosphorylation, of phosphorylase. The hexadecameric enzyme, with subunit composition (alpha,beta,gamma,delta)4, serves to integrate hormonal, metabolic and neural signals, leading to the breakdown of glycogen stores in muscle and liver. Each subunit of PhK is encoded by a separate gene, and multiple tissue specific isoforms exist for all of the subunits. While much work has been done to elucidate the biochemical regulation of PhK catalysis, little information regarding transcriptional regulation of this complex oligomer is available. Our work seeks to elucidate those elements responsible for the temporal and spatial regulation of PhK gene expression by providing for the identification and functional analysis of all human alpha, beta and gamma promoters. The proposed project has four aims directed at this goal. 1) The putative PhK promoter reg ions for all alpha, beta and gamma isoforms will be identified through bioinformatic analysis of the human genome database and amplified by PCR. 2) The minimal sequence necessary to convey spatial and temporal gene expression will be determined for each promoter. 3) Key regulatory cis- and trans-acting factors will be identified for each promoter. 4) Microarray analysis will be performed on RNA from PhK deficient mice to identify novel proteins involved in coordinated gene expression. Knowledge gained from these aims will help determine how PhK expression is orchestrated to ensure that metabolic plasticity is modulated in response to changing energy demands.

该子项目是利用NIH/NCRR资助的中心赠款提供的资源的许多研究子项目之一。子项目和研究者（PI）可能从另一个NIH来源获得主要资金，因此可以在其他CRISP条目中表示。所列机构为中心，不一定是研究者所在机构。磷酸化酶激酶（PhK）是参与糖原分解途径的关键调节酶，通过磷酸化催化磷酸化酶的活化。具有亚基组成（α、β、γ、δ）4的十六聚体酶用于整合激素、代谢和神经信号，导致肌肉和肝脏中糖原储存的分解。PhK的每个亚基由单独的基因编码，并且所有亚基都存在多种组织特异性同种型。虽然已经做了很多工作来阐明PhK催化的生化调节，但关于这种复杂寡聚体的转录调节的信息很少。我们的工作旨在阐明负责的时间和空间调控的PhK基因的表达，通过提供所有人类α，β和γ启动子的识别和功能分析的元素。针对这一目标，拟议的项目有四个目标。1)所有α、β和γ亚型的推定PhK启动子区域将通过人类基因组数据库的生物信息学分析鉴定，并通过PCR扩增。2)对于每个启动子，将确定传达空间和时间基因表达所需的最小序列。3)将确定每个启动子的关键调控顺式和反式作用因子。4)将对PhK缺陷小鼠的RNA进行微阵列分析，以鉴定参与协调基因表达的新型蛋白质。从这些目标中获得的知识将有助于确定PhK表达是如何协调的，以确保代谢可塑性响应于不断变化的能量需求而被调节。