Analysis of a Borrelia burgdorferi phosphodiesterase demonstrates a role for cyclic-di-guanosine monophosphate in motility and virulence.
Analysis of a Borrelia burgdorferi phosphodiesterase demonstrates a role for cyclic-di-guanosine monophosphate in motility and virulence.
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DOI：
10.1111/j.1365-2958.2010.07191.x
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发表时间：
2010-07-01
影响因子：
3.6
通讯作者：
Motaleb MA
中科院分区：
文献类型：
--
作者：
Sultan SZ;Pitzer JE;Miller MR;Motaleb MA
伯氏疏螺旋体的基因组编码了一组被认为参与循环二gmp代谢的基因。虽然BB0419被证明是一种二胍酸环化酶，但BB0419或任何假定的环化二gmp代谢基因对伯氏伯氏杆菌运动和发病机制的影响程度尚未报道。在这里，我们鉴定和表征磷酸二酯酶（BB0363）。BB0363特异性水解环二gmp， Km为0.054 μM，证实其为功能性环二gmp磷酸二酯酶。利用新开发的不影响下游基因表达的无启动子抗生素盒构建了bb0363的靶向突变。突变细胞表现出游泳模式的改变，表明循环二gmp在调节伯氏疏螺旋体运动中的作用。此外，bb0363突变细胞在小鼠体内不具有传染性，表明循环二gmp在伯氏疏螺旋体感染中的重要作用。在食用naïve小鼠的血液后，突变细胞能够在肩胛骨蜱体内存活；然而，感染突变细胞的蜱不能感染naïve小鼠。运动和感染表型在基因互补后恢复。这些结果揭示了环二gmp、伯氏疏螺旋体运动与莱姆病发病机制之间的重要联系。提出了环二gmp影响运动和感染的机制。
The genome of Borrelia burgdorferi encodes a set of genes putatively involved in cyclic-di-GMP metabolism. Although BB0419 was shown to be a diguanylate cyclase, the extent to which bb0419 or any of the putative cyclic-di-GMP metabolizing genes impact B. burgdorferi motility and pathogenesis has not yet been reported. Here we identify and characterize a phosphodiesterase (BB0363). BB0363 specifically hydrolyzed cyclic-di-GMP with a Km of 0.054 μM, confirming it is a functional cyclic-di-GMP phosphodiesterase. A targeted mutation in bb0363 was constructed using a newly developed promoterless antibiotic cassette that does not affect downstream gene expression. The mutant cells exhibited an altered swimming pattern, indicating a function for cyclic-di-GMP in regulating B. burgdorferi motility. Furthermore, the bb0363 mutant cells were not infectious in mice, demonstrating an important role for cyclic-di-GMP in B. burgdorferi infection. The mutant cells were able to survive within Ixodes scapularis ticks after a blood meal from naïve mice; however, ticks infected with the mutant cells were not able to infect naïve mice. Both motility and infection phenotypes were restored upon genetic complementation. These results reveal an important connection between cyclic-di-GMP, B. burgdorferi motility and Lyme disease pathogenesis. A mechanism by which cyclic-di-GMP influences motility and infection is proposed.
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影响因子：
4.5
作者：
Girgis, Hany S.;Liu, Yirchung;Ryu, William S.;Tavazoie, Saeed
通讯作者：
Tavazoie, Saeed
影响因子：
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作者：
Amikam, D;Galperin, MY
通讯作者：
Galperin, MY
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3.2
作者：
Beyhan, Sinem;Tischler, Anna D.;Yildiz, Fitnat H.
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Yildiz, Fitnat H.
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作者：
Boehm, Alex;Kaiser, Matthias;Jenal, Urs
通讯作者：
Jenal, Urs
影响因子：
2.9
作者：
Chang, AL;Tuckerman, JR;Gilles-Gonzalez, MA
通讯作者：
Gilles-Gonzalez, MA