Biological and Biochemical Roles of Two Distinct Cyclic Dimeric Adenosine 3',5'-Monophosphate- Associated Phosphodiesterases in Streptococcus mutans.
Biological and Biochemical Roles of Two Distinct Cyclic Dimeric Adenosine 3',5'-Monophosphate- Associated Phosphodiesterases in Streptococcus mutans.
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DOI：
10.3389/fmicb.2018.02347
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发表时间：
2018
影响因子：
5.2
通讯作者：
Hasegawa Y
中科院分区：
文献类型：
--
作者：
Konno H;Yoshida Y;Nagano K;Takebe J;Hasegawa Y
环二聚腺苷3′，5 ′-单磷酸（c-di-AMP）是一种新近发现的细菌第二信使，在细菌生物膜形成等过程中发挥重要作用。由二腺苷酸环化酶酶促产生，并由c-di-AMP磷酸二酯酶裂解。本研究对变形链球菌（Streptococcus mutans）两种不同的磷酸二酯酶GdpP（SMU_2140c）和DhhP（SMU_1297）的生物化学和生物学作用进行了研究。液相色谱-串联质谱（LC-MS-MS）分析表明，c-di-AMP被截短型重组GdpP降解为磷酸腺苷酰腺苷（pApA），pApA被重组DhhP裂解为AMP。缺失dhhP基因（ΔdhhP）以及缺失gdpP和dhhP基因（ΔgdpPΔdhhP）的框内缺失突变体显示出比野生型和缺失gdpP基因的突变菌株（ΔgdpP; p < 0.01）显著更多的生物膜形成。此外，在与dhhP互补后，生物膜形成恢复到野生型菌株的水平。光学和电子显微镜观察显示ΔdhhP和ΔgdpPΔdhhP突变体自聚集成大的细胞团，与增加的生物膜形成相关，但在野生型、ΔgdpP或补充有gdpP和dhhP的菌株的培养物中未观察到细胞团。因此，dhhP的缺失可能导致细菌细胞聚集体的形成和随后生物膜产生的增加。
Cyclic dimeric adenosine 3′,5′-monophosphate (c-di-AMP), a recently identified secondary messenger in bacteria, plays a role in several bacterial processes, including biofilm formation. It is enzymatically produced by diadenylate cyclase and cleaved by c-di-AMP phosphodiesterase. c-di-AMP is believed to be essential for the viability of bacterial cells that produce it. In the current study, the biochemical and biological roles of GdpP (SMU_2140c) and DhhP (SMU_1297), two distinct Streptococcus mutans phosphodiesterases involved in the pathway producing AMP from c-di-AMP, were investigated. Liquid chromatography-tandem mass spectrometry revealed that c-di-AMP was degraded to phosphoadenylyl adenosine (pApA) by truncated recombinant GdpP, and pApA was cleaved by recombinant DhhP to yield AMP. In-frame deletion mutants lacking the dhhP gene (ΔdhhP) and both the gdpP and dhhP genes (ΔgdpPΔdhhP) displayed significantly more biofilm formation than the wild-type and a mutant strain lacking the gdpP gene (ΔgdpP; p < 0.01). Furthermore, biofilm formation was restored to the level of the wild type strain upon complementation with dhhP. Optical and electron microscopy observations revealed that ΔdhhP and ΔgdpPΔdhhP mutants self-aggregated into large cell clumps, correlated with increased biofilm formation, but cell clumps were not observed in cultures of wild-type, ΔgdpP, or strains complemented with gdpP and dhhP. Thus, deletion of dhhP presumably leads to the formation of bacterial cell aggregates and a subsequent increase in biofilm production.
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