NosP Signaling Modulates the NO/H-NOX-Mediated Multicomponent c-Di-GMP Network and Biofilm Formation in Shewanella oneidensis
NosP Signaling Modulates the NO/H-NOX-Mediated Multicomponent c-Di-GMP Network and Biofilm Formation in Shewanella oneidensis
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DOI：
10.1021/acs.biochem.9b00706
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发表时间：
2019-12-03
期刊：
影响因子：
2.9
通讯作者：
Boon, Elizabeth M.
中科院分区：
文献类型：
--
作者：
Nisbett, Lisa-Marie;Binnenkade, Lucas;Boon, Elizabeth M.
当细菌在自我分泌的胞外多糖基质中聚集时，形成生物膜;它们对抗生素具有抗性并与疾病有关。已知一氧化氮（NO）通过连接到H-NOX（血红素-NO/氧结合）结构域来介导许多细菌中的生物膜形成。然而，大多数NO响应细菌缺乏含H-NOX结构域的蛋白质。我们已经确定了另一个NO敏感蛋白（NosP），这是预测参与双组分信号和生物膜调节在许多物种。在这里，我们证明了NosP参与了先前描述的H-NOX/NO响应多组分c-di-GMP信号网络希瓦氏菌oneidensis。缺乏nosP或其共顺反子激酶nahK（先前的hnoS）的菌株产生不成熟的生物膜，而hnoX和hnoK（响应于NO/H-NOX的激酶）突变体导致野生型生物膜结构。我们证明，NosP调节NahK以及HnoK的自磷酸化活性。HnoK和NahK已经显示出调节三种反应调节物（HnoB、HnoC和HnoD），它们一起构成NO响应性多组分c-di-GMP信号传导网络。在这里，我们提出NosP/NahK增加了H-NOX/HnoK的调节，以调节这种c-di-GMP信号网络，并最终通过HnoK和NahK控制磷酸盐的流量来形成生物膜。此外，似乎NosP和H-NOX在推拉机制中相互抵消; NosP/NahK通过抑制H-NOX/HnoK信号传导促进生物膜形成，这本身降低了生物膜形成的程度。NO的加入导致c-di-GMP和生物膜形成的减少，主要是通过HnoK活性的去抑制。
Biofilms form when bacteria aggregate in a self-secreted exopolysaccharide matrix; they are resistant to antibiotics and implicated in disease. Nitric oxide (NO) is known to mediate biofilm formation in many bacteria via ligation to H-NOX (heme-NO/oxygen binding) domains. Most NO-responsive bacteria, however, lack H-NOX domain-containing proteins. We have identified another NO-sensing protein (NosP), which is predicted to be involved in two-component signaling and biofilm regulation in many species. Here, we demonstrate that NosP participates in the previously described H-NOX/NO-responsive multicomponent c-di-GMP signaling network in Shewanella oneidensis. Strains lacking either nosP or its co-cistronic kinase nahK (previously hnoS) produce immature biofilms, while hnoX and hnoK (kinase responsive to NO/H-NOX) mutants result in wild-type biofilm architecture. We demonstrate that NosP regulates the autophosphorylation activity of NahK as well as HnoK. HnoK and NahK have been shown to regulate three response regulators (HnoB, HnoC, and HnoD) that together comprise a NO-responsive multicomponent c-di-GMP signaling network. Here, we propose that NosP/NahK adds regulation on top of H-NOX/HnoK to modulate this c-di-GMP signaling network, and ultimately biofilm formation, by governing the flux of phosphate through both HnoK and NahK. In addition, it appears that NosP and H-NOX act to counter each other in a push-pull mechanism; NosP/NahK promotes biofilm formation through inhibition of H-NOX/HnoK signaling, which itself reduces the extent of biofilm formation. Addition of NO results in a reduction of c-di-GMP and biofilm formation, primarily through disinhibition of HnoK activity.