Studies of a Bacterial Protein N-Lysine Acetyltransferase

描述（由申请人提供）：关于n -赖氨酸乙酰化在蛋白质功能中的作用的拟议工作对所有研究中心代谢和基因表达调控的生物学家具有潜在的广泛意义，因为它与癌症和人类衰老有关。我们有一个优秀的模型原核生物（肠沙门氏菌），其中执行所提出的工作。研究这种细菌的主要优点是我们能够研究不同遗传背景下感兴趣的突变的影响，从中我们可以了解乙酰化/去乙酰化系统有效所需的辅助功能。利用肠球菌复杂的遗传系统将极大地促进具有所需功能的菌株的工程设计。这种模式生物对于生产和分离修饰或未修饰的蛋白质也非常有价值，这些蛋白质的功能可以在分离中详细研究。蛋白质乙酰化与中心代谢之间的联系已经在细菌、小鼠和人类细胞中建立起来。因此，肠球菌的研究结果可能会使我们对所有生命形式的代谢途径整合的理解取得重要进展。对细菌系统的进一步了解将反过来促进生产低分子量化合物、抗生素和其他次生代谢物、肽、酶和蛋白质的生理策略的实施，这些对农业和制药工业以及一般的生物技术都有价值。从这些研究中获得的关于乙酰化蛋白质的酶的功能作为细胞中乙酰辅酶a水平的功能的新知识将揭示酶的功能如何在响应细胞生理状态的变化或响应改变辅酶a稳态的环境刺激时被调节。这一新信息将有助于设计和构建对特定刺激反应改变的乙酰转移酶的变体形式。这些变异蛋白可能对具有理想代谢能力的菌株的基因工程至关重要。

DESCRIPTION (provided by applicant): The proposed work on the role of N-Lysine acetylation on protein function has potentially broad implications of interest to all biologists studying central metabolism and regulation of gene expression as it pertains to cancer and human aging. We have an excellent model prokaryotic organism (Salmonella enterica) in which to perform the proposed work. The chief advantage of working with this bacterium is our ability to study the effect of mutations of interest in diverse genetic backgrounds from which we can learn about accessory functions needed for the acetylation/deacetylation system to be efficient. Use of the S. enterica sophisticated genetic system will greatly facilitate the engineering of strains with required functionalities. This model organism is also extremely valuable for the production and isolation of modified or unmodified proteins whose function can be studied in detail in isolation. The connection between protein acetylation and central metabolism has been established in bacteria, murine and human cells. Thus, results from studies in S. enterica will likely lead to important advances in our understanding of metabolic pathway integration in all forms of life. Improved understanding of the bacterial system will in turn facilitate the implementation of physiological strategies for the production of low molecular weight compounds, antibiotics and other secondary metabolites, peptides, enzymes, and proteins of value to the agricultural and pharmaceutical industries, and biotechnology in general. New knowledge obtained from these studies about the function of the enzyme that acetylates proteins as a function of acetyl-CoA levels in the cell will shed light on how enzyme function might be regulated in response to changes in the physiological state of the cell or in response to environmental stimuli that alter CoA homeostasis. This new information will be useful in the design and construction of variant forms of the acetyltransferase with altered response to specific stimuli. These variant proteins may be critical in the genetic engineering of strains with desirable metabolic capabilities.