Dissertation Research: Pathway Analysis of Ethanol and Acetic Acid Detoxification in Drosophila

乙醇和乙酸耐受性在D.黑腹果蝇和果蝇的种类。 与乙醇和乙酸解毒途径中的不同步骤相关的性状的非独立性强调了对乙醇和乙酸耐受性的途径分析的必要性。 这一完整途径的分析受到阻碍的事实，只有ADH和醛氧化酶（ALDOX）可以在生物化学和分子水平上进行研究，离开乙醇解毒途径的第三步，由乙酰辅酶A合成酶（AcCoAS）催化，uncharacterized。 从D.黑腹果蝇和最近开发的AcCoAS反应的分子探针现在允许乙醇和乙酸耐受性的完整途径分析。 我们假设，这两种耐受性的选择对整个解毒途径起作用，并将塑造该途径中三种关键酶之间的变异和共变异模式。 将采用技术的组合来定量核苷酸序列数据、mRNA表达水平酶活性和对乙醇和乙酸的耐受性。由该博士论文改进资助的研究采用进化定量遗传方法来表征遗传变异和酶活性变异之间的关系，以及途径中所有三个步骤的后果。这种对乙醇和乙酸耐受性的变化。 这些发现将在代谢控制理论的背景下进行分析，并使我们能够预测生物体性能选择的遗传和生化反应

0105085ClarkEthanol and acetic acid tolerance are highly positively correlated across populations of D. melanogaster and species of Drosophila. The non-independence of traits associated with different steps in the ethanol and acetic acid detoxification pathway underscores the need for a pathway analysis of ethanol and acetic acid tolerance. Analysis of this complete pathway was hindered by the fact that only ADH and aldehyde oxidase (ALDOX) could be studied at the biochemical and molecular levels, leaving the third step in the ethanol detoxification pathway, catalyzed by acetyl Co-A synthetase (AcCoAS), uncharacterized. Identification of AcCoAS from the complete genome sequence of D. melanogaster and recently developed molecular probes for the AcCoAS reaction now allow for a complete pathway analysis of ethanol and acetic acid tolerance. We hypothesize that selection for both tolerances operates on the entire detoxification pathway and will shape the patterns of variation and covariation between the three key enzymes in the pathway. A combination of techniques will be employed to quantify variation in and covariation between nucleotide sequence data, mRNA expression levels, enzyme activities and tolerance to both ethanol and acetic acid.The research funded by this Doctoral Dissertation Improvement Grant takes an evolutionary quantitative genetic approach to characterize the relationship between genetic variation and enzyme activity variation at all three steps in the pathway and the consequences of this variation on ethanol and acetic acid tolerance. The findings will be analyzed in the context of metabolic control theory and will allow us to make predictions about the genetic and biochemical responses to selection on organismal performance