Studies of a Bacterial Protein N-Lysine Acetyltransferase

描述(由申请人提供)：关于N-赖氨酸乙酰化对蛋白质功能的作用的拟议工作具有潜在的广泛意义，所有研究与癌症和人类衰老有关的中枢代谢和基因表达调控的生物学家都对此感兴趣。我们有一个很好的原核生物模型(肠沙门氏菌)来执行拟议的工作。与这种细菌合作的主要优势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/脱乙酰化系统高效所需的辅助功能。使用肠杆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。这种模式生物对于生产和分离修饰或未修饰的蛋白质也是非常有价值的，这些蛋白质的功能可以在分离中详细研究。已经在细菌、小鼠和人类细胞中建立了蛋白质乙酰化和中枢代谢之间的联系。因此，对肠链球菌的研究结果可能会导致我们对所有生命形式中代谢途径整合的理解取得重要进展。对细菌系统的更好的理解将反过来促进生产低分子化合物、抗生素和其他次级代谢物、多肽、酶和蛋白质的生理学策略的实施，这些对农业和制药工业以及整个生物技术都有价值。从这些研究中获得的关于乙酰化蛋白质的酶的功能与细胞中乙酰辅酶A水平的函数关系的新知识，将有助于阐明如何调节酶功能以响应细胞生理状态的变化或响应改变辅酶A动态平衡的环境刺激。这一新信息将有助于设计和构建对特定刺激具有不同反应的不同形式的乙酰基转移酶。这些变异蛋白在具有理想代谢能力的菌株的基因工程中可能是关键的。

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.