Center for Oral Health and Systemic Disease, School of Dentistry, University of Louisville Louisville, KY, USA.
Front Cell Infect Microbiol. 2012 Jun 12;2:79. doi: 10.3389/fcimb.2012.00079. eCollection 2012.
Porphyromonas gingivalis is a major etiological agent in chronic and aggressive forms of periodontal disease. The organism is an asaccharolytic anaerobe and is a constituent of mixed species biofilms in a variety of microenvironments in the oral cavity. P. gingivalis expresses a range of virulence factors over which it exerts tight control. High-throughput sequencing technologies provide the opportunity to relate functional genomics to basic biology. In this study we report qualitative and quantitative RNA-Seq analysis of the transcriptome of P. gingivalis. We have also applied RNA-Seq to the transcriptome of a ΔluxS mutant of P. gingivalis deficient in AI-2-mediated bacterial communication. The transcriptome analysis confirmed the expression of all predicted ORFs for strain ATCC 33277, including 854 hypothetical proteins, and allowed the identification of hitherto unknown transcriptional units. Twelve non-coding RNAs were identified, including 11 small RNAs and one cobalamin riboswitch. Fifty-seven genes were differentially regulated in the LuxS mutant. Addition of exogenous synthetic 4,5-dihydroxy-2,3-pentanedione (DPD, AI-2 precursor) to the ΔluxS mutant culture complemented expression of a subset of genes, indicating that LuxS is involved in both AI-2 signaling and non-signaling dependent systems in P. gingivalis. This work provides an important dataset for future study of P. gingivalis pathophysiology and further defines the LuxS regulon in this oral pathogen.
牙龈卟啉单胞菌是慢性和侵袭性牙周病的主要病因。该生物体是一种无碳源的厌氧菌,是口腔内各种微环境中混合物种生物膜的组成部分。牙龈卟啉单胞菌表达一系列毒力因子,并对其进行严格控制。高通量测序技术为将功能基因组学与基础生物学联系起来提供了机会。在这项研究中,我们报告了牙龈卟啉单胞菌转录组的定性和定量 RNA-Seq 分析。我们还将 RNA-Seq 应用于牙龈卟啉单胞菌ΔluxS 突变体的转录组分析,该突变体缺乏 AI-2 介导的细菌通讯。转录组分析证实了 ATCC 33277 菌株所有预测的 ORF 的表达,包括 854 个假设蛋白,并确定了以前未知的转录单元。鉴定了 12 个非编码 RNA,包括 11 个小 RNA 和一个钴胺素核糖开关。在 LuxS 突变体中,有 57 个基因的表达发生了差异调节。向ΔluxS 突变体培养物中添加外源性合成 4,5-二羟基-2,3-戊二酮(DPD,AI-2 前体)可补充部分基因的表达,表明 LuxS 参与了牙龈卟啉单胞菌的 AI-2 信号转导和非信号转导系统。这项工作为未来研究牙龈卟啉单胞菌的病理生理学提供了重要的数据集,并进一步定义了该口腔病原体中 LuxS 调节子。