Effects of extracellular DNA and DNA-binding protein on the development of a Streptococcus intermedius biofilm
Effects of extracellular DNA and DNA-binding protein on the development of a Streptococcus intermedius biofilm
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DOI：
10.1111/jam.12202
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发表时间：
2013-07-01
影响因子：
4
通讯作者：
Miyake, Y.
中科院分区：
文献类型：
--
作者：
Nur, A.;Hirota, K.;Miyake, Y.
目的研究同源和异源胞外DNA（eDNAs）和组蛋白样DNA结合蛋白（HLP）对中间链球菌生物膜形成和刚性的影响。方法与结果用0中心点1%结晶紫染色法测定生物被膜质量，扫描电镜观察。分别用7-羟基-9H-（1，3-二氯-9，9-二甲基吖啶-2-酮）和抗HLP抗体染色，不固定法检测eDNA和细胞外HLP（eHLP）在生物膜中的定位。DNase I处理（200 U ml ~（-1））显著降低了生物膜的形成和细胞密度。证实了eHLP和eDNA在生物膜中的共定位。向中间链球菌培养物中添加从中间链球菌、其他革兰氏阳性细菌、革兰氏阴性细菌或人KB细胞纯化的eDNA（高达lg ml-1）增加了中间链球菌的所有测试菌株、野生型、HLP下调菌株和对照菌株的生物膜质量。相比之下，添加eDNA（>1g ml-1）降低了所有中间链球菌菌株的生物膜质量。结论eDNA和eHLP在生物被膜的形成和刚性中起重要作用。eDNA和HLP靶向策略可能适用于细菌生物膜相关感染性疾病的新型治疗。
Aims The aim of this study was to clarify the effects of homologous and heterologous extracellular DNAs (eDNAs) and histone-like DNA-binding protein (HLP) on Streptococcus intermedius biofilm development and rigidity. Methods and Results Formed biofilm mass was measured with 0 center dot 1% crystal violet staining method and observed with a scanning electron microscope. The localizations of eDNA and extracellular HLP (eHLP) in formed biofilm were detected by staining with 7-hydoxyl-9H-(1,3-dichloro-9,9-dimethylacridin-2-one) and anti-HLP antibody without fixation, respectively. DNase I treatment (200U ml-1) markedly decreased biofilm formation and cell density in biofilms. Colocalization of eHLP and eDNA in biofilm was confirmed. The addition of eDNA (up to 1g ml-1) purified from Strep.intermedius, other Gram-positive bacteria, Gram-negative bacteria, or human KB cells into the Strep.intermedius culture increased the biofilm mass of all tested strains of Strep.intermedius, wild-type, HLP-downregulated strain and control strains. In contrast, the addition of eDNA (>1g ml-1) decreased the biofilm mass of all Strep.intermedius strains. Conclusions These findings demonstrated that eDNA and eHLP play crucial roles in biofilm development and its rigidity. Significance and Impact of the Study eDNA- and HLP-targeting strategies may be applicable to novel treatments for bacterial biofilm-related infectious diseases.