Mechanisms of Glucose Signal Transduction in Yeast

酵母中葡萄糖信号转导机制

基本信息

批准号：
7254131
负责人：
Martin C Schmidt
金额：
$ 30.2万
依托单位：
UNIVERSITY OF PITTSBURGH AT PITTSBURGH
依托单位国家：
美国
项目类别：
财政年份：
1991
资助国家：
美国
起止时间：
1991-07-01 至 2008-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7254131
关键词：
5&apos -AMP-activated protein kinase Alanine Alleles Binding Biochemical Genetics Biological Assay C-terminal Cells Client Co-Immunoprecipitations Complex Condition Discrimination Drug usage Enzymes Event GLC2 protein Gene Deletion Glucose Grant Homologous Gene In Vitro Length Mammalian Cell Maps Mass Spectrum Analysis Measures Messenger RNA Metformin Molecular Chaperones Molecular Conformation Mutagenesis Non-Insulin-Dependent Diabetes Mellitus Northern Blotting Numbers Nutrient Pathway interactions Phosphoric Monoester Hydrolases Phosphorylation Phosphorylation Site Phosphotransferases Play Post-Transcriptional Regulation Protein Kinase Proteins Regulation Research Personnel Role Role playing therapy Saccharomyces cerevisiae Saccharomycetales Signal Pathway Signal Transduction Site Stress Testing Ubiquitin Western Blotting Yeasts genetic analysis genetic regulatory protein in vivo member mutant novel programs protein protein interaction response upstream kinase

项目摘要

DESCRIPTION (provided by applicant): The Snf1 protein kinase plays a central role in nutrient stress signaling in the budding yeast, Saccharomyces cerevisiae. The mammalian homologue of Snf1, the AMP-activated protein kinase (AMPK) plays an equally important role in mammalian cells. Recently AMPK was shown to be the target of metformin, the most widely prescribed drug used to treat type 2 diabetes. We propose to study the regulation of this medically important enzyme in yeast. In the previous grant period we developed an assay for the phosphorylation state of the Snf1 activation loop. We showed that phosphorylation of Snf1 was a critical step for the activation of the Snf1 kinase and that the phosphorylation of Snf1 was catalyzed by a distinct upstream kinase. Recently, we were able to demonstrate that there are three upstream kinases (Pakl, Tos3 and Elm1) that are capable of activating Snf1 in vivo. The identification of the Snf1 activating kinases was highly significant because it immediately led to the identification of the mammalian kinase that activates AMPK. In the next grant period we propose to continue our studies of the regulation of the Snf1 kinase signaling pathway. We will focus on upstream events in this pathway and determine the mechanism that regulate the activity of the three Snf1-activating kinases. We will determine whether there are distinct forms of the Snf1 kinase complex that are specialized for response to different stresses. We will assess the regulatory significance of phosphorylation sites in the Snf1 kinase complex that we have mapped by mass spectrometry. Finally, biochemical and genetic analyses have identified a number of candidates for regulators of the Snf1 kinase signaling pathway. These candidates proteins will be studied to determine if they associate with and regulate the activity of the Snf1 kinase.

描述（由申请人提供）：SNF1蛋白激酶在发芽的酵母中的营养应激信号传导中起着核心作用，酿酒酵母。 SNF1的哺乳动物同源物，AMP激活的蛋白激酶（AMPK）在哺乳动物细胞中起同样重要的作用。最近，AMPK被证明是二甲双胍的靶标，二甲双胍是用于治疗2型糖尿病的最广泛规定的药物。我们建议研究对酵母中这种重要的重要酶的调节。在上一个赠款期间，我们开发了SNF1激活环的磷酸化状态的测定法。我们表明，SNF1的磷酸化是激活SNF1激酶的关键步骤，并且SNF1的磷酸化被独特的上游激酶催化。最近，我们能够证明能够在体内激活SNF1的三种上游激酶（PAKL，TOS3和ELM1）。 SNF1激活激酶的鉴定非常重要，因为它立即导致鉴定激活AMPK的哺乳动物激酶。在下一个赠款期间，我们建议继续研究SNF1激酶信号通路的调节。我们将重点关注该途径中的上游事件，并确定调节三种SNF1激活激酶活性的机制。我们将确定SNF1激酶复合物的不同形式是否专门针对不同的压力。我们将评估SNF1激酶复合物中磷酸化位点的调节意义，该磷酸化位点通过质谱法绘制了。最后，生化和遗传分析已经确定了SNF1激酶信号通路调节剂的许多候选物。这些候选蛋白将进行研究，以确定它们是否与SNF1激酶的活性相关联。