Novel Mechanisms of Beta-lactam Resistance in Staph Aureus

金黄色葡萄球菌β-内酰胺耐药的新机制

• 批准号: 10318974
• 负责人: Som Chatterjee
• 金额: $ 65.37万
• 依托单位: UNIVERSITY OF MARYLAND BALTIMORE
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-08-01 至 2024-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10318974
• 关键词: Affect; Affinity; Ampicillin; Antibiotics; Attenuated; Bacteria; Binding; Biochemical; Biological Assay; Biology; Cell Wall; Cell physiology; Cell surface; Cells; Chemicals; Clinical; Cytolysins; Data; Defect; Follow-Up Studies; Future; Gene Expression; Genetic; Genetic Transcription; Genus staphylococcus; Goals; Human; Investigation; Knowledge; Mediating; Mediator; Methicillin Resistance; Missense Mutation; Molecular Weight; Morbidity - disease rate; Mutation; Mutation Detection; Nature; Penicillin-Binding Proteins; Penicillins; Peptidyltransferase; Periodicity; Pharmaceutical Preparations; Phenotype; Physiological; Process; Production; Promoter Regions; Proteins; Proteomics; RNA; Regulation; Research; Resistance; Role; Second Messenger Systems; Signal Transduction; Staphylococcus aureus; Treatment Failure; Virulence; Virulence Factors; Yin-Yang; beta-Lactam Resistance; beta-Lactamase; beta-Lactams; cost; crosslink; experimental study; genome sequencing; in vivo; inhibitor; lead candidate; loss of function mutation; mortality; novel; novel therapeutic intervention; overexpression; pathogen; pathogenic bacteria; phosphoric diester hydrolase; promoter; resistance mechanism; resistant strain; screening; surveillance study; whole genome

Abstract We have identified a novel mode of high-level, broad-spectrum β-lactam resistance in S. aureus that is not mediated by PBP2a, the penicillin-binding protein (PBP) that confers methicillin resistance. PBP4, a non- essential PBP, and GdpP, the only known phosphodiesterase (PDE) that mediates cyclic-di-adenosine-mono- phosphate (CDA) degradation, have critical roles in this type of resistance. Mutations that enhance PBP4's ability to make a highly cross-linked bacterial cell wall and loss-of-function mutations in GdpP are the genetic basis responsible for this uncanonical resistance. The highly cross-linked cell wall formation is driven either independently or cooperatively by two distinct biochemical features of PBP4, a) structural changes in its protein due to missense mutations and b) its overexpression due to mutations in its promoter region. The loss-of-function mutations in GdpP result in elevated concentrations of CDA in bacterial cells. CDA is a newly discovered cell-signaling second messenger in bacteria which acts as an allosteric regulator by binding to its effectors (proteins and RNAs). CDA broadly affects gene expression and controls GdpP related β-lactam resistant phenotypes in a concentration dependent manner, suggesting that it is the deterministic factor in resistance. However, the precise role(s) of CDA in mediating β-lactam resistance as well as other vital processes of S. aureus is currently unknown. These functional alterations of PBP4 and GdpP likely come at the cost of bacterial virulence due to depletion of cell wall associated proteins and attenuated production of cytolysins, respectively. This indicates a unique yin-yang relationship between two key pathogenic factors of S. aureus, β-lactam resistance and virulence. We will investigate the fundamental basis of the functional changes in PBP4 and GdpP that lead to resistance and their impact on bacterial virulence. Aim 1: To determine the mechanism of PBP4-mediated β-lactam resistance and the role of PBP4 in cell wall composition. The relative contribution of PBP4 missense and promoter mutations on cell wall synthesis will be evaluated biochemically and structurally. The mechanism(s) that control pbp4 expression will be investigated to identify regulator(s) and to determine how they confer PBP4-mediated β-lactam resistance. PBP4's role on bacterial cell surface associated virulence factor expression will be determined. Aim 2: To define the role of cyclic-di-adenosine-mono-phosphate (CDA) signaling in S. aureus. Genetic and chemical proteomic approaches will be taken to identify CDA mediator/s in the bacteria that are responsible for β-lactam resistance and virulence defect. Finally, our preliminary data suggest the presence of a novel CDA specific phosphodiesterase in S. aureus besides GdpP. We will identify this novel phosphodiesterase. The proposed research will advance knowledge of basic cellular processes in S. aureus.

抽象的 我们已经确定了一种新型的金黄色葡萄球菌中高级，广谱β-内酰胺的抗性 由PBP2A介导的，是赋予甲氧西林耐药性的青霉素结合蛋白（PBP）。 pbp4，非 - 必需PBP和GDPP，这是唯一介导环状 - 二腺苷 - 单核 - 单核酶的已知磷酸二酯酶（PDE） 磷酸盐（CDA）降解，在这种抗性中具有关键作用。增强PBP4的突变 具有高度交联细菌细胞壁和GDPP功能丧失突变的能力是遗传 对这种非正式的抵抗负责的基础。高度交联的细胞壁形成驱动 通过PBP4的两个不同的生化特征独立或合作，a）其蛋白质的结构变化 由于错义突变和b）由于其启动子区域的突变引起的过表达。 GDPP的功能丧失突变导致细菌细胞中CDA浓度升高。 CDA 是细菌中新发现的细胞信号第二信使，用作变构调节剂 与其作用（蛋白质和RNA）结合。 CDA广泛影响基因表达并控制GDPP相关 以浓度依赖性方式β-内酰胺抗性表型，表明它是确定性的 电阻的因素。但是，CDA在介导β-内酰胺耐药性以及其他方面的精度作用 金黄色葡萄球菌的重要过程目前尚不清楚。 PBP4和GDPP的这些功能改变可能是由于细菌病毒而造成的 细胞壁相关蛋白的消耗和细胞蛋白酶的产生分别消耗。这表明 金黄色葡萄球菌，β-内酰胺抗性和 病毒。我们将研究PBP4和GDPP功能变化的基本基础 抗性及其对细菌病毒的影响。目标1：确定PBP4介导的机制 β-内酰胺抗性和PBP4在细胞壁组成中的作用。 PBP4的相对贡献 细胞壁合成上的错义和启动子突变将在生化和结构上进行评估。这 将研究控制PBP4表达的机制，以识别调节剂并确定如何 他们会遇到PBP4介导的β-内酰胺耐药性。 PBP4在细菌细胞表面相关病毒中的作用 因子表达将确定。目标2：定义环状二腺苷 - 单磷酸的作用 （CDA）金黄色葡萄球菌中的信号传导。将采用遗传和化学蛋白质组学方法来鉴定CDA 细菌中的介体导致β-内酰胺耐药性和病毒缺陷。最后，我们的 初步数据表明，除GDPP以外，在金黄色葡萄球菌中存在新型CDA特异性磷酸二酯酶。 我们将确定这种新型的磷酸二酯酶。拟议的研究将提高基本细胞的知识 金黄色葡萄球菌中的过程。

项目成果

Ceftaroline-Resistant, Daptomycin-Tolerant, and Heterogeneous Vancomycin-Intermediate Methicillin-Resistant Staphylococcus aureus Causing Infective Endocarditis.

头孢洛林耐药、达托霉素耐受和异种万古霉素中间甲氧西林耐药金黄色葡萄球菌引起感染性心内膜炎。

• DOI: 10.1128/aac.01235-16
• 发表时间: 2017
• 期刊: Antimicrobial agents and chemotherapy
• 影响因子: 4.9
• 作者: Nigo,Masayuki;Diaz,Lorena;Carvajal,LinaP;Tran,TrucT;Rios,Rafael;Panesso,Diana;Garavito,JuanD;Miller,WilliamR;Wanger,Audrey;Weinstock,George;Munita,JoseM;Arias,CesarA;Chambers,HenryF
• 通讯作者: Chambers,HenryF

In Vivo Detection of Cyclic-di-AMP in Staphylococcus aureus.

• DOI: 10.1021/acsomega.2c04538
• 发表时间: 2022-09-13
• 期刊: ACS OMEGA
• 影响因子: 4.1
• 作者: Mukkayyan, Nagaraja;Poon, Raymond;Sander, Philipp N.;Lai, Li-Yin;Zubair-Nizami, Zahra;Hammond, Ming C.;Chatterjee, Som S.
• 通讯作者: Chatterjee, Som S.

PBP4-mediated β-lactam resistance among clinical strains of Staphylococcus aureus.

金黄色葡萄球菌临床菌株中 PBP4 介导的 β-内酰胺耐药性。

• DOI: 10.1093/jac/dkab201
• 发表时间: 2021
• 期刊: The Journal of antimicrobial chemotherapy
• 作者: Satishkumar,Nidhi;Alexander,JAndrewN;Poon,Raymond;Buggeln,Emma;Argudín,MariaA;Strynadka,NatalieCJ;Chatterjee,SomS
• 通讯作者: Chatterjee,SomS

A Nonclassical Mechanism of β-Lactam Resistance in Methicillin-Resistant Staphylococcus aureus and Its Effect on Virulence.

• DOI: 10.1128/spectrum.02284-22
• 发表时间: 2022-12-21
• 期刊: Microbiology spectrum
• 影响因子: 3.7

Structural basis of broad-spectrum β-lactam resistance in Staphylococcus aureus.

• DOI: 10.1038/s41586-022-05583-3
• 发表时间: 2023-01
• 期刊: NATURE
• 影响因子: 64.8
• 作者: Alexander, J. Andrew N.;Worrall, Liam J.;Hu, Jinhong;Vuckovic, Marija;Satishkumar, Nidhi;Poon, Raymond;Sobhanifar, Solmaz;Rosell, Federico I.;Jenkins, Joshua;Chiang, Daniel;Mosimann, Wesley A.;Chambers, Henry F.;Paetzel, Mark;Chatterjee, Som S.;Strynadka, Natalie C. J.
• 通讯作者: Strynadka, Natalie C. J.