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对研究中心代谢和基因表达调控的所有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。我们有一个很好的模型原核生物（沙门氏菌），在其中进行拟议的工作。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中感兴趣的突变的影响，从中我们可以了解乙酰化/脱乙酰化系统有效所需的辅助功能。使用S。enterica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化。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的情况下详细研究。蛋白质乙酰化和中枢代谢之间的联系已经在细菌、鼠和人细胞中建立。因此，S.肠道可能会导致我们对所有形式生命中代谢途径整合的理解的重要进展。对细菌系统的更好理解反过来将促进用于生产对农业和制药工业有价值的低分子量化合物、抗生素和其他次级代谢物、肽、酶和蛋白质以及一般生物技术的生理策略的实施。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水平的函数的新知识将揭示酶功能如何响应于细胞生理状态的变化或响应于改变辅酶A稳态的环境刺激而被调节。这一新的信息将是有用的，在设计和建设的变异形式的乙酰转移酶与改变响应特定的刺激。这些变异蛋白质在具有所需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.