Establishment of cell polarity by Par-1 kinase during Drosophila oogenesis: assessment of biochemically identified Par-1 substrates as targets in vivo

果蝇卵子发生过程中 Par-1 激酶建立细胞极性：评估生化鉴定的 Par-1 底物作为体内靶标

• 批准号: 5331590
• 负责人: Dr. Anne Ephrussi, Ph.D.
• 金额: --
• 依托单位: Europäisches Laboratorium für Molekularbiologie (EMBL)
• 依托单位国家: 德国
• 项目类别: Priority Programmes
• 财政年份: 2001
• 资助国家: 德国
• 起止时间: 2000-12-31 至 2008-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/5331590?language=en
• 关键词: Establishment; cell; polarity; Par; kinase

The gene par-1 encodes an evolutionarily conserved Ser/Thr kinase. Genetic data show that par-1 is required for the establishment of polarity in organisms as divergent as worms and flies. Although biochemical data suggest a function of the mammalian Par-1 homologue, MARK, in regulating cytoskeletal organisation by phosphorylating microtubule-associated proteins (MAPs), this property of the kinase is not sufficient to explain the polarity defects observed in the par-1 mutants. We use Drosophila oogenesis as a model to understand the mechanisms of cell polarization. par-1 is required several times for establishment and maintenance of cell polarity during oogenesis, but the mechanism by which Par-1 regulates cell polarity is not understood, as neither in C. elegans nor in Drosophila are Par-1 target proteins known. We will use "in vitro expression cloning" to identify cDNAs encoding proteins that are phosphorylated by Par-1. Pools of cDNAs will be transcribed and translated in vitro to be tested as substrates of the kinase. We will systematically screen the Drosophila genome for Par-1 targets.

par1基因编码一种进化上保守的丝氨酸/苏氨酸激酶。遗传数据显示，par-1是蠕虫和苍蝇等生物极性形成所必需的。尽管生化数据表明哺乳动物Par-1同源物MARK通过磷酸化微管相关蛋白（MAPs）来调节细胞骨架组织，但该激酶的这一特性不足以解释在Par-1突变体中观察到的极性缺陷。我们使用果蝇卵发生作为模型来理解细胞极化的机制。在卵子发生过程中，par-1需要多次建立和维持细胞极性，但par-1调节细胞极性的机制尚不清楚，因为在秀丽隐杆线虫和果蝇中都不知道par-1靶蛋白。我们将使用“体外表达克隆”来鉴定编码par1磷酸化蛋白的cdna。大量的cdna将在体外转录和翻译，作为激酶的底物进行测试。我们将系统地筛选果蝇基因组中的Par-1靶点。