Investigation of the physiological significance of protein acetylation in Bacillus subtilis

枯草芽孢杆菌蛋白质乙酰化生理意义的研究

• 批准号: 10247780
• 负责人: Valerie Jean Carabetta
• 金额: $ 35.78万
• 依托单位: ROWAN UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-09-01 至 2025-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10247780
• 关键词: Acetylation; Acetyltransferase; Address; Affect; Bacillus subtilis; Bacteria; Bacterial Physiology; Bacterial Proteins; Biochemical; Biological Process; Biology; Cell division; Cell physiology; Chromosome Structures; Chromosomes; Code; DNA; Deacetylation; Development; Drug Targeting; Drug Tolerance; Enzymes; Eukaryota; Foundations; Gene Expression; Genetic Recombination; Goals; Histone Acetylation; Histone Code; Histones; Investigation; Knowledge; Lysine; Mass Spectrum Analysis; Modeling; Modification; N-terminal; Phase; Physiological; Post-Translational Protein Processing; Process; Protein Acetylation; Protein Family; Proteins; Proteomics; Regulation; Research; Site; Structure; Tail; Techniques; Transact; Work; antimicrobial; antimicrobial drug; base; design; inhibitor/antagonist; member; novel; programs; repaired

ABSTRACT Until recently, N-lysine acetylation was thought to be rare in bacteria, but is now appreciated to affect hundreds of bacterial proteins with diverse cellular functions. Acetylation was initially discovered as a post-translational modification (PTM) on the unstructured, highly basic N-terminal tails of eukaryotic histones. Histone acetylation constitutes part of the “histone code,” and regulates chromosome compaction and various DNA processes, such as gene expression, replication, repair and recombination. In eukaryotes, acetylation regulates many other proteins in addition to histones, involved in a wide array of important biological processes. This observation is also true in bacteria, as evidenced by the characterization of the acetylomes of more than 30 different bacterial species. However, the physiological significance of the vast majority of these modifications remains unknown. In addition, the mechanisms of acetylation and deacetylation, and the bacterial enzymes involved are not completely understood. To address these gaps in knowledge, we have focused on studying the acetylation of the essential, histone-like protein HBsu in Bacillus subtilis. In bacteria, the nucleoid is compacted and organized by the action of nucleoid-associated proteins (NAPs). HBsu is a member of the most widely conserved NAP family, and is considered a functional equivalent of eukaryotic histones. We found that HBsu contains seven novel acetylation sites, and this raised the exciting possibility that these modifications represent a “histone-like” code in bacteria. So far, we discovered that acetylation of HBsu at key lysine residues is required to maintain normal chromosome compaction. Additionally, we identified the second protein acetyltransferase in B. subtilis. The overall goal of our research program is to decipher this code. Our recent progress supports the hypotheses that acetylation of HBsu regulates cell division and sporulation, and that there are additional enzymes involved in regulating acetylation. The short- term goals of this work are to define the enzymatic mechanism of regulation of HBsu acetylation and determine the significance of HBsu acetylation in the regulation of DNA transactions, stationary phase development and drug tolerance. Additionally, we will develop new biochemical and mass-spectrometry based proteomics techniques for the study of acetylation in bacteria. Our long-term goals are to characterize additional HBsu PTMs, identify and characterize novel enzymes of acetylation, and perform a detailed structural and biochemical analysis with acetylated HBsu and novel enzymes. Ultimately, we will design novel inhibitors of bacterial acetylation enzymes or acetylated HBsu and assess their efficacy as potential novel antimicrobial therapies. Together, these studies may demonstrate the existence of a histone- like code in bacteria, an unexpected and exciting new field of biology. Furthermore, these studies will provide the foundation for designing novel antimicrobial drugs that target protein acetylation, either the enzymes or key acetylated targets.

摘要 直到最近，N-赖氨酸乙酰化被认为在细菌中是罕见的，但现在被理解为影响数百种细菌。 具有多种细胞功能的细菌蛋白质。乙酰化最初被发现是一种翻译后修饰 (PTM)非结构化的，高度碱性的真核组蛋白的N-末端尾部。组蛋白乙酰化构成了 “组蛋白密码”，并调节染色体压缩和各种DNA过程，如基因表达，复制， 修复和重组。在真核生物中，乙酰化除了调节组蛋白外，还调节许多其他蛋白质，参与了蛋白质的合成。 一系列重要的生物过程。这一观察结果在细菌中也是正确的，正如表征所证明的那样。 超过30种不同细菌的乙酰体。然而，绝大多数的生理意义 这些修改仍然未知。此外，还对乙酰化和脱乙酰化反应的机理以及 所涉及的细菌酶还不完全清楚。为了解决这些知识差距，我们重点关注 研究枯草芽孢杆菌中必需的组蛋白样蛋白HBsu的乙酰化。在细菌中， 通过类核蛋白（nucleotide associated proteins，NAPs）的作用而紧密组织。HBSU是世界上最广泛的 保守的NAP家族，并且被认为是真核组蛋白的功能等同物。我们发现HBsu含有 七个新的乙酰化位点，这提出了令人兴奋的可能性，这些修饰代表了“组蛋白样” 细菌中的密码。到目前为止，我们发现HBsu在关键赖氨酸残基上的乙酰化是维持正常的 染色体致密化此外，我们还鉴定了B中的第二个蛋白乙酰转移酶。枯草杆菌。总目标 就是破译这段代码我们最近的进展支持了HBsu乙酰化 调节细胞分裂和孢子形成，并且存在参与调节乙酰化的另外的酶。短- 这项工作的长期目标是确定HBsu乙酰化调节的酶促机制，并确定HBsu乙酰化酶的活性。 HBsu乙酰化在DNA转录调控、稳定期形成和药物耐受中的重要性。 此外，我们将开发新的生物化学和质谱为基础的蛋白质组学技术的研究， 细菌中的乙酰化作用。我们的长期目标是表征其他HBsu PTM，识别和表征新的 酶的乙酰化，并进行详细的结构和生化分析与乙酰化HBsu和新的 内切酶最终，我们将设计新型的细菌乙酰化酶或乙酰化HBsu抑制剂，并评估它们的作用。 作为潜在的新型抗微生物疗法的功效。总之，这些研究可以证明组蛋白的存在- 就像细菌中的密码一样，这是一个意想不到的令人兴奋的生物学新领域。此外，这些研究将提供 为设计靶向蛋白质乙酰化的新型抗微生物药物奠定了基础，无论是酶还是关键的乙酰化酶， 目标的