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.

生物膜是附着在各种表面上的固定细菌群落。生物膜的存在通常会使细菌对有效对抗其自由生活（浮游）对应物的清除策略产生抵抗力。因此，了解促进固定生活方式和不受束缚生活方式之间转换的分子线索可以指导我们在有益和有害的情况下对生物膜形成的反应。该项目试图在系统水平上表征伴随生物膜形成的细胞变化，以及在分子水平上特定的传感器蛋白家族在生物膜形成/溶解中的作用。学生培养是这项工作的重点。学生将得到具有多种技术专业知识的科学家团队的共同指导，并将有机会在国家实验室进行变革性研究。还将为研究生开设一个蛋白质组学研讨会，以培训下一代科学家掌握这一尖端技术。科学目标是使用先进的结构和蛋白质组学方法来定义与跨多个物种的生物膜形成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）感知氧化剂和/或一氧化氮（NO）的机制，从而导致生物膜形成或溶解。将利用 X 射线晶体学和核磁共振 (NMR) 以及 X 射线吸收精细结构 (EXAFS) 进行结构测定。高级蛋白质组学和磷酸蛋白质组学将使用 H-NOX 缺失突变体在生物体水平上详细介绍这一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及其如何进化以满足不同细菌物种的需求产生详细的机制理解。该奖项反映了 NSF 的法定使命，并通过使用基金会的智力优势和更广泛的评估进行评估，被认为值得支持。影响审查标准。