Department of Biological Science and Technology, Institute of Technology and Science, The University of Tokushima Graduate School, 2 chome, Minami-josanjima, Tokushima 770-8506, Japan.
Cell Stress Chaperones. 2012 Jan;17(1):41-55. doi: 10.1007/s12192-011-0284-4. Epub 2011 Aug 6.
Streptococcus intermedius is a facultatively anaerobic, opportunistic pathogen that causes purulent infections and abscess formation. The DnaK chaperone system has been characterized in several pathogenic bacteria and seems to have important functions in stress resistance and pathogenicity. However, the role of DnaK in S. intermedius remains unclear. Therefore, we constructed a dnaK knockout mutant that exhibited slow growth, thermosensitivity, accumulation of GroEL in the cell, and reduced cytotoxicity to HepG2 cells. The level of secretion of a major pathogenic factor, intermedilysin, was not affected by dnaK mutation. We further examined the function and property of the S. intermedius DnaK chaperone system by using Escherichia coli ΔdnaK and ΔrpoH mutant strains. S. intermedius DnaK could not complement the thermosensitivity of E. coli ΔdnaK mutant. However, the intact S. intermedius DnaK chaperone system could complement the thermosensitivity and acid sensitivity of E. coli ΔdnaK mutant. The S. intermedius DnaK chaperone system could regulate the activity and stability of the heat shock transcription factor σ(32) in E. coli, although S. intermedius does not utilize σ(32) for heat shock transcription. The S. intermedius DnaK chaperone system was also able to efficiently eliminate the aggregated proteins from ΔrpoH mutant cells. Overall, our data showed that the S. intermedius DnaK chaperone system has important functions in quality control of cellular proteins but has less participation in the modulation of expression of pathogenic factors.
中间链球菌是一种兼性厌氧、机会性病原体,可引起化脓性感染和脓肿形成。DnaK 伴侣系统已在几种致病菌中得到了描述,似乎在应激抗性和致病性方面具有重要功能。然而,DnaK 在中间链球菌中的作用尚不清楚。因此,我们构建了一个 dnaK 敲除突变体,该突变体表现出生长缓慢、热敏性、细胞内 GroEL 积累和对 HepG2 细胞的细胞毒性降低。主要致病因子中间溶素的分泌水平不受 dnaK 突变的影响。我们进一步使用大肠杆菌 ΔdnaK 和 ΔrpoH 突变株研究了中间链球菌 DnaK 伴侣系统的功能和特性。中间链球菌 DnaK 不能补充大肠杆菌 ΔdnaK 突变体的热敏性。然而,完整的中间链球菌 DnaK 伴侣系统可以补充大肠杆菌 ΔdnaK 突变体的热敏性和酸性敏感性。中间链球菌 DnaK 伴侣系统可以调节大肠杆菌中热休克转录因子 σ(32)的活性和稳定性,尽管中间链球菌不利用 σ(32)进行热休克转录。中间链球菌 DnaK 伴侣系统还能够有效地从 ΔrpoH 突变细胞中清除聚集的蛋白质。总的来说,我们的数据表明,中间链球菌 DnaK 伴侣系统在细胞蛋白质的质量控制中具有重要功能,但在调节致病因子的表达方面参与较少。