Collaborative Research: H-NOX homologues in biofilm formation: A combined molecular and systems level approach.

合作研究：生物膜形成中的 H-NOX 同系物：分子和系统水平相结合的方法。

• 批准号: 2103677
• 负责人: Kelly Chacon
• 金额: $ 28.14万
• 依托单位: Reed College
• 依托单位国家: 美国
• 项目类别: Continuing Grant
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-07-01 至 2025-06-30
• 项目状态: 未结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=2103677&HistoricalAwards=false
• 关键词: Collaborative; Research; NOX; homologues; biofilm

Biofilms are stationary communities of bacteria that adhere to various surfaces. Inhabitation of a biofilm often makes bacteria resistant to clearance strategies that are effective against their free-living (planktonic) counterparts. As such, understanding the molecular cues that promote switching between the stationary and untethered lifestyles can guide our responses to biofilm formation in both beneficial and detrimental contexts. This project seeks to characterize cellular changes accompanying biofilm formation at the systems level, as well as the role of a particular family of sensor proteins in biofilm formation/dissolution at the molecular level. Student training is a major emphasis of this work. Students will be co-mentored by a team of scientists with expertise in diverse techniques and will have the opportunity to conduct transformative research at national laboratories. A proteomics workshop for graduate students will also be developed to train the next generation of scientists in this cutting-edge technology. The scientific objective is to define conserved and novel mechanisms of bacterial signaling relevant to biofilm formation across multiple species using advanced structural and proteomics methods. At the molecular level, this will include determining the mechanisms by which the heme nitric oxide/oxygen binding proteins (H-NOX) sense oxidants and/or nitric oxide (NO), resulting in biofilm formation or dissolution. Structure determination using X-ray crystallography and nuclear magnetic resonance (NMR) will be utilized as well as X-ray absorption fine-structure (EXAFS). Advanced proteomics and phosphoproteomics will detail the signaling networks involved in this process at the organismal level using H-NOX deletion mutants. This combination of approaches will yield detailed mechanistic understanding of an important signaling process and how it has evolved to meet the needs of diverse bacterial species.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.

生物膜是粘附在各种表面的细菌固定群落。生物膜的居住通常会使细菌抗清除策略具有有效的自由生活（浮游生物）对应物。因此，了解促进固定和不受束缚生活方式之间切换的分子提示可以指导我们在有益和有害环境中对生物膜形成的反应。该项目旨在表征系统级别伴随生物膜形成的细胞变化，以及特定的传感器蛋白家族在生物膜形成/分子水平的溶解中的作用。学生培训是这项工作的主要重点。学生将由具有多种技术专业知识的科学家团队共同征收，并将有机会在国家实验室进行变革性研究。还将开发针对研究生的蛋白质组学研讨会，以培训这项尖端技术的下一代科学家。科学目标是使用先进的结构和蛋白质组学方法来定义与多种物种相关的细菌信号传导的保守和新颖机制。在分子水平上，这将包括确定血红素一氧化氮/氧结合蛋白（H-NOX）含义氧化剂和/或一氧化氮（NO）的机制，从而导致生物膜形成或溶解。使用X射线晶体学和核磁共振（NMR）的结构测定以及X射线吸收精细结构（EXAFS）。先进的蛋白质组学和磷蛋白质组学将使用H-NOX缺失突变体在生物水平上详细介绍此过程中涉及的信号网络。这种方法的结合将对重要的信号传导过程产生详细的机械理解，以及它如何发展以满足各种细菌物种的需求。该奖项反映了NSF的法定任务，并被认为是通过基金会的智力优点和更广泛的评估而被视为值得支持的。影响审查标准。