Probing the drivers of Staphylococcus aureus biofilm protein amyloidogenesis and disrupting biofilms with engineered protein disaggregases.
Probing the drivers of Staphylococcus aureus biofilm protein amyloidogenesis and disrupting biofilms with engineered protein disaggregases.
复制标题
作者：
酚溶性调节素（psm）是金黄色葡萄球菌生物膜的主要蛋白质成分。居住在生物膜的保护环境中，细菌可以迅速进化并获得抗菌素耐药性，这可能导致持续感染，如耐甲氧西林金黄色葡萄球菌（MRSA）引起的感染。在它们的可溶性形式中，psm阻碍宿主的免疫反应，并可以增加MRSA的毒力潜力。psm也自组装成不溶性的功能性淀粉样蛋白，有助于生物膜的结构支架。PSM肽在生物膜中的具体作用仍然知之甚少。在这里，我们报道了一种遗传可处理的酵母模型系统的发展，用于研究PSMα肽的特性。酵母中PSMα肽的表达驱动了采用囊泡样结构的毒性不溶性聚集体的形成。利用该系统，我们探索了PSMα聚集的分子驱动因素，以描绘PSM之间的关键异同，并确定了驱动PSM特征的关键残基。生物膜是一个主要的公共卫生威胁；因此，破坏生物膜是一个关键目标。为了溶解由多种淀粉样蛋白和淀粉样蛋白组成的聚集体，我们开发了Hsp104的工程变体，这是一种来自酵母的六聚体AAA+蛋白分解酶。在这里，我们证明了增强的Hsp104变体对抗PSMα肽的毒性和聚集。此外，我们证明了增强的Hsp104变体可以驱动预形成的金黄色葡萄球菌生物膜的分解。我们认为，这种新的酵母模型可以作为一个强大的平台，用于筛选破坏PSM聚集的药物，Hsp104分解气体可能是一种有前途的工具，用于安全的酶破坏生物膜。生物膜是由细菌分泌的复杂混合物，形成细菌可以嵌入的物质。这一过程改变了细菌的特性，使它们对清除变得更有抵抗力，这可能产生多重耐药菌株，如耐甲氧西林金黄色葡萄球菌（MRSA）。在这里，我们研究了酚溶性调节素（psm），这是金黄色葡萄球菌分泌的淀粉样蛋白，可以整合到生物膜中。生物膜的研究具有挑战性，因此我们开发了一种新的遗传可处理酵母模型来研究psm。我们使用我们的系统来了解psm的几个关键特性。我们还证明了淀粉样蛋白分解酶Hsp104的变体可以破坏psm，更重要的是，可以溶解预先形成的金黄色葡萄球菌生物膜。我们认为该系统可以作为一种强大的筛选工具，并且Hsp104分解气体可能是探索生物膜破坏剂的新途径。
Phenol-soluble modulins (PSMs) are the primary proteinaceous component of Staphylococcus aureus biofilms. Residence in the protective environment of biofilms allows bacteria to rapidly evolve and acquire antimicrobial resistance, which can lead to persistent infections such as those caused by methicillin-resistant S. aureus (MRSA). In their soluble form, PSMs hinder the immune response of the host and can increase the virulence potential of MRSA. PSMs also self-assemble into insoluble functional amyloids that contribute to the structural scaffold of biofilms. The specific roles of PSM peptides in biofilms remain poorly understood. Here, we report the development of a genetically tractable yeast model system for studying the properties of PSMα peptides. Expression of PSMα peptides in yeast drives the formation of toxic insoluble aggregates that adopt vesicle-like structures. Using this system, we probed the molecular drivers of PSMα aggregation to delineate key similarities and differences among the PSMs and identified a crucial residue that drives PSM features. Biofilms are a major public health threat; thus, biofilm disruption is a key goal. To solubilize aggregates comprised of a diverse range of amyloid and amyloid-like species, we have developed engineered variants of Hsp104, a hexameric AAA+ protein disaggregase from yeast. Here, we demonstrate that potentiated Hsp104 variants counter the toxicity and aggregation of PSMα peptides. Further, we demonstrate that a potentiated Hsp104 variant can drive the disassembly of preformed S. aureus biofilms. We suggest that this new yeast model can be a powerful platform for screening for agents that disrupt PSM aggregation and that Hsp104 disaggregases could be a promising tool for the safe enzymatic disruption of biofilms. Biofilms are complex mixtures secreted by bacteria that form a material in which the bacteria can become embedded. This process transforms the properties of the bacteria, and they become more resistant to removal, which can give rise to multidrug-resistant strains, such as methicillin-resistant Staphylococcus aureus (MRSA). Here, we study phenol-soluble modulins (PSMs), which are amyloidogenic proteins secreted by S. aureus, that become incorporated into biofilms. Biofilms are challenging to study, so we have developed a new genetically tractable yeast model to study the PSMs. We used our system to learn about several key features of the PSMs. We also demonstrate that variants of an amyloid disaggregase, Hsp104, can disrupt the PSMs and, more importantly, dissolve preformed S. aureus biofilms. We propose that our system can be a powerful screening tool and that Hsp104 disaggregases may be a new avenue to explore for biofilm disruption agents.
登录
查看更多内容
影响因子：
9.8
作者：
Ju S;Tardiff DF;Han H;Divya K;Zhong Q;Maquat LE;Bosco DA;Hayward LJ;Brown RH Jr;Lindquist S;Ringe D;Petsko GA
通讯作者：
Petsko GA
影响因子：
64.5
作者：
DeSantis ME;Leung EH;Sweeny EA;Jackrel ME;Cushman-Nick M;Neuhaus-Follini A;Vashist S;Sochor MA;Knight MN;Shorter J
通讯作者：
Shorter J
影响因子：
--
作者：
Castellano LM;Bart SM;Holmes VM;Weissman D;Shorter J
通讯作者：
Shorter J
影响因子：
4.3
作者：
Jackrel, Meredith E.;Shorter, James
通讯作者：
Shorter, James
DOI：
10.1126/science.1245296
发表时间：
2013-11-22
期刊：
Science (New York, N.Y.)
影响因子：
--
作者：
Chung CY;Khurana V;Auluck PK;Tardiff DF;Mazzulli JR;Soldner F;Baru V;Lou Y;Freyzon Y;Cho S;Mungenast AE;Muffat J;Mitalipova M;Pluth MD;Jui NT;Schüle B;Lippard SJ;Tsai LH;Krainc D;Buchwald SL;Jaenisch R;Lindquist S
通讯作者：
Lindquist S