Kuboniwa Masae, Hendrickson Erik L, Xia Qiangwei, Wang Tiansong, Xie Hua, Hackett Murray, Lamont Richard J
Department of Preventive Dentistry, Osaka University Graduate School of Dentistry, Osaka, Japan.
BMC Microbiol. 2009 May 19;9:98. doi: 10.1186/1471-2180-9-98.
Porphyromonas gingivalis is a periodontal pathogen that resides in a complex multispecies microbial biofilm community known as dental plaque. Confocal laser scanning microscopy showed that P. gingivalis can assemble into communities in vitro with Streptococcus gordonii and Fusobacterium nucleatum, common constituents of dental plaque. Whole cell quantitative proteomics, along with mutant construction and analysis, were conducted to investigate how P. gingivalis adapts to this three species community.
1156 P. gingivalis proteins were detected qualitatively during comparison of the three species model community with P. gingivalis incubated alone under the same conditions. Integration of spectral counting and summed signal intensity analyses of the dataset showed that 403 proteins were down-regulated and 89 proteins up-regulated. The proteomics results were inspected manually and an ontology analysis conducted using DAVID. Significant decreases were seen in proteins involved in cell shape and the formation of the cell envelope, as well as thiamine, cobalamin, and pyrimidine synthesis and DNA repair. An overall increase was seen in proteins involved in protein synthesis. HmuR, a TonB dependent outer membrane receptor, was up-regulated in the community and an hmuR deficient mutant was deficient in three species community formation, but was unimpaired in its ability to form mono- or dual-species biofilms.
Collectively, these results indicate that P. gingivalis can assemble into a heterotypic community with F. nucleatum and S. gordonii, and that a community lifestyle provides physiologic support for P. gingivalis. Proteins such as HmuR, that are up-regulated, can be necessary for community structure.
牙龈卟啉单胞菌是一种牙周病原体,存在于称为牙菌斑的复杂多物种微生物生物膜群落中。共聚焦激光扫描显微镜显示,牙龈卟啉单胞菌可在体外与牙菌斑的常见成分戈登链球菌和具核梭杆菌组装成群落。进行了全细胞定量蛋白质组学以及突变体构建和分析,以研究牙龈卟啉单胞菌如何适应这种三物种群落。
在相同条件下将三物种模型群落与单独培养的牙龈卟啉单胞菌进行比较时,定性检测到1156种牙龈卟啉单胞菌蛋白质。对数据集的光谱计数和总信号强度分析的整合表明,403种蛋白质下调,89种蛋白质上调。手动检查蛋白质组学结果,并使用DAVID进行本体分析。参与细胞形状和细胞壁形成的蛋白质以及硫胺素、钴胺素和嘧啶合成及DNA修复的蛋白质显著减少。参与蛋白质合成的蛋白质总体上有所增加。HmuR是一种依赖TonB的外膜受体,在群落中上调,hmuR缺陷突变体在三物种群落形成中存在缺陷,但在形成单物种或双物种生物膜的能力方面未受损。
总体而言,这些结果表明牙龈卟啉单胞菌可与具核梭杆菌和戈登链球菌组装成异型群落,并且群落生活方式为牙龈卟啉单胞菌提供了生理支持。上调的蛋白质如HmuR可能是群落结构所必需的。