Inactivation of Cyclic Di-GMP Binding Protein TDE0214 Affects the Motility, Biofilm Formation, and Virulence of Treponema denticola
Inactivation of Cyclic Di-GMP Binding Protein TDE0214 Affects the Motility, Biofilm Formation, and Virulence of Treponema denticola
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DOI：
10.1128/jb.00610-13
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发表时间：
2013-09-01
影响因子：
3.2
通讯作者：
Li, Chunhao
中科院分区：
文献类型：
--
作者：
Bian, Jiang;Liu, Xiangyang;Li, Chunhao
环二聚GMP（c-di-GMP）作为一种普遍存在的第二信使，在许多细菌中被研究。口腔螺旋体齿垢密螺旋体（Treponema denticola）是与人类牙周炎相关的牙周病原体，具有复杂的c-di-GMP信号网络。然而，它的功能仍然未被探索。在这份报告中，PilZ样c-di-GMP结合蛋白（TDE 0214）进行了研究，以调查c-di-GMP在螺旋体中的作用。TDE 0214具有PilZ结构域，其具有两个特征基序：RXXXR和DXSXXG。生物化学研究表明，TDE 0214以特异性方式结合c-di-GMP，解离常数（K-d）值为1.73 μ M，与其他报道的c-di-GMP结合蛋白相比处于较低范围。探讨c-di-GMP在T.构建了TDE 0214缺失突变体（Td Delta 214），并对其进行了详细的分析。首先，游泳板和单细胞跟踪分析表明，Td Delta 214具有异常的游泳行为：突变体比野生型运动性更低，逆转频率更高。其次，我们发现Td Delta 214的生物膜形成基本上被抑制（类似于6.0倍减少）。最后，使用小鼠皮肤脓肿模型的体内研究显示，Td Delta 214的侵袭性和诱导皮肤脓肿和宿主体液免疫应答的能力显著减弱，表明TDE 0214对T.齿垢总的来说，这里报告的结果表明TDE 0214在螺旋体的运动性、生物膜形成和毒力中起重要作用。本研究也为进一步揭示c-di-GMP信号网络在T.齿垢
As a ubiquitous second messenger, cyclic dimeric GMP (c-di-GMP) has been studied in numerous bacteria. The oral spirochete Treponema denticola, a periodontal pathogen associated with human periodontitis, has a complex c-di-GMP signaling network. However, its function remains unexplored. In this report, a PilZ-like c-di-GMP binding protein (TDE0214) was studied to investigate the role of c-di-GMP in the spirochete. TDE0214 harbors a PilZ domain with two signature motifs: RXXXR and DXSXXG. Biochemical studies showed that TDE0214 binds c-di-GMP in a specific manner, with a dissociation constant (K-d) value of 1.73 mu M, which is in the low range compared to those of other reported c-di-GMP binding proteins. To reveal the role of c-di-GMP in T. denticola, a TDE0214 deletion mutant (Td Delta 214) was constructed and analyzed in detail. First, swim plate and single-cell tracking analyses showed that Td Delta 214 had abnormal swimming behaviors: the mutant was less motile and reversed more frequently than the wild type. Second, we found that biofilm formation of Td Delta 214 was substantially repressed (similar to 6.0-fold reduction). Finally, in vivo studies using a mouse skin abscess model revealed that the invasiveness and ability to induce skin abscesses and host humoral immune responses were significantly attenuated in Td Delta 214, indicative of the impact that TDE0214 has on the virulence of T. denticola. Collectively, the results reported here indicate that TDE0214 plays important roles in motility, biofilm formation, and virulence of the spirochete. This report also paves a way to further unveil the roles of the c-di-GMP signaling network in the biology and pathogenicity of T. denticola.