Rice Kelly C, Mann Ethan E, Endres Jennifer L, Weiss Elizabeth C, Cassat James E, Smeltzer Mark S, Bayles Kenneth W
Department of Pathology and Microbiology, University of Nebraska Medical Center, Omaha, NE 68198, USA.
Proc Natl Acad Sci U S A. 2007 May 8;104(19):8113-8. doi: 10.1073/pnas.0610226104. Epub 2007 Apr 23.
The Staphylococcus aureus cidA and lrgA genes have been shown to affect cell lysis under a variety of conditions during planktonic growth. It is hypothesized that these genes encode holins and antiholins, respectively, and may serve as molecular control elements of bacterial cell lysis. To examine the biological role of cell death and lysis, we studied the impact of the cidA mutation on biofilm development. Interestingly, this mutation had a dramatic impact on biofilm morphology and adherence. The cidA mutant (KB1050) biofilm exhibited a rougher appearance compared with the parental strain (UAMS-1) and was less adherent. Propidium iodide staining revealed that KB1050 accumulated more dead cells within the biofilm population relative to UAMS-1, indicative of reduced cell lysis. In agreement with this finding, quantitative real-time PCR experiments demonstrated the presence of 5-fold less genomic DNA in the KB1050 biofilm relative to UAMS-1. Furthermore, treatment of the UAMS-1 biofilm with DNase I caused extensive cell detachment, whereas similar treatment of the KB1050 biofilm had only a modest effect. These results demonstrate that cidA-controlled cell lysis plays a significant role during biofilm development and that released genomic DNA is an important structural component of S. aureus biofilm.
金黄色葡萄球菌的cidA和lrgA基因已被证明在浮游生长的各种条件下会影响细胞裂解。据推测,这些基因分别编码孔蛋白和抗孔蛋白,可能作为细菌细胞裂解的分子控制元件。为了研究细胞死亡和裂解的生物学作用,我们研究了cidA突变对生物膜形成的影响。有趣的是,这种突变对生物膜形态和粘附性有显著影响。与亲本菌株(UAMS-1)相比,cidA突变体(KB1050)生物膜外观更粗糙,粘附性更低。碘化丙啶染色显示,相对于UAMS-1,KB1050在生物膜群体中积累了更多的死细胞,表明细胞裂解减少。与此发现一致,定量实时PCR实验表明,相对于UAMS-1,KB1050生物膜中的基因组DNA含量少5倍。此外,用DNase I处理UAMS-1生物膜会导致大量细胞脱落,而对KB1050生物膜进行类似处理只有适度的效果。这些结果表明,cidA控制的细胞裂解在生物膜形成过程中起重要作用,释放的基因组DNA是金黄色葡萄球菌生物膜的重要结构成分。