AGE AND DISEASE ASSOCIATED CHANGES IN CREATINE KINASE

本研究的长期目标是了解 能量转导的基因调控，特别是在高表达的组织中， 能量需求，如心脏、大脑和平滑肌。 肌酸 激酶（CK）基因家族在能量过程中具有核心作用 转导，并由四个独立的基因，这是高度 以组织特异性、适应性和发育模式进行调节。 的 这四个基因的功能由磷酸肌酸（CP） 穿梭假说，其中线粒体ATP是底物， 线粒体肌酸激酶（MtCK），可逆地催化 将高能磷酸盐转化为肌酸。 CP穿梭机需要 协调调节细胞溶质和线粒体Cks，以及 强调了MtCK在调节氧化应激中的关键地位， 磷酸化 这项建议的重点是调节能量转导， 老化的人体组织 选择CK基因进行研究是因为 缺乏关于能量基因调控的信息 衰老的人体组织中的线粒体转导和生物发生， 特别是大脑，因为初步数据表明， CK基因、BCK和普遍存在的MtCK（uMtCK）的表达可能是 由其保守的3 '-非翻译区进行转录后调节 （UTR）。 具体的目标将确定所有的协调调节 四个CK基因在老化的人体组织，并测试假设，3 '- KD基因的UTR是保守的，因为它们发挥转录后作用， 控制表达。 这些具体目标是：1）确定CK 在人脑、心脏和小肠中的表达和活性， 老年人和年轻人的尸检组织。 2)识别和表征 使用RNA凝胶迁移率与CK mRNA的3 '-UTR结合的蛋白质- RNA结合蛋白的转移测定和亲和纯化。 明确能量传递的基因调控过程， 大大提高了我们对能源重要性的认识 在健康、衰老和疾病的非肌节组织中产生， 例如中风或衰老。

The long-term objective of this research is to understand the process of gene regulation of energy transduction, particularly in tissues with high energy requirements such as heart, brain, and smooth muscle. The Creatine Kinase (CK) gene family has a central role in the process of energy transduction, and is composed of four separate genes which are highly regulated in a tissue-specific, adaptive, and developmental pattern. The function of these four genes is described by the Creatine Phosphate (CP) shuttle hypothesis, in which mitochondrial ATP is a substrate for mitochondrial creatine kinase (MtCK) which reversibly catalyzes the transfer of high energy phosphate to creatine. The CP shuttle requires coordinate regulation of the cytosolic and mitochondrial Cks, and emphasizes the key position that MtCK has in regulating oxidative phosphorylation. The focus of this proposal is the regulation of energy transduction in aging human tissues. The CK genes have been chosen for study because of the paucity of information regarding gene regulation of energy transduction and mitochondrial biogenesis in aging human tissues, especially brain, and because preliminary data suggest that coordinate expression of the CK genes, BCK and ubiquitous MtCK (uMtCK) may be regulated posttranscriptionally by their conserved 3'-untranslated regions (UTR). The specific aims will define the coordinate regulation of all four CK genes in aging human tissues, and test the hypothesis that the 3'- UTRs of the KD genes are conserved because they exert posttranscriptional control of expression. These specific aims are: 1) Determine CK expression and activity in human brain, heart, and small intestine using autopsy tissues from aged and young adults. 2) Identify and characterize proteins binding to the 3'-UTRs of the CK mRNAs using RNA gel mobility- shift assays and affinity purification of RNA-binding proteins. Defining the process of gene regulation of energy transduction will significantly advance our understanding of the importance of energy production in non-sarcomeric tissues in health, aging, and in disease, such as stroke or senility.