Institute of Cytology, Russian Academy of Sciences, St. Petersburg, Russia.
J Cell Biochem. 2013 Jul;114(7):1568-74. doi: 10.1002/jcb.24498.
Serratia grimesii are non-pathogenic bacteria capable, however, to invade eukaryotic cells provided that they synthesize intracellular metalloprotease grimelysin (Bozhokina et al. [2011] Cell. Biol. Int. 35: 111-118). To elucidate how invasion of grimelysin containing bacteria depends on physiological state of host cells, we studied the effect of N-acetylcysteine (NAC) on susceptibility of HeLa cells to invasion by the wild-type S. grimesii and recombinant E. coli expressing grimelysin gene. Incubation of HeLa cells with 10 mM NAC resulted in changes of cell morphology and disassembly of actin cytoskeleton that were reversed when NAC was removed from the culture medium. Both in the presence of NAC and upon its removal, the entry of grimelysin producing bacteria increased by a factor of 1.5-2 and 3-3.5 for wild-type S. grimesii and recombinant E. coli, respectively. This effect does not correlate with cytoskeleton rearrangements but may be due to the NAC-induced up-regulation of cell surface receptors playing a role in cell adhesion and cell-cell junctions. A twofold difference in the efficiency of S. grimesii and recombinant E. coli to enter the NAC-treated cells suggests that the entry of the wild-type and recombinant bacteria occurs via different receptors which activity is differently affected by NAC.
粘质沙雷氏菌是非致病性细菌,但能够合成细胞内金属蛋白酶格里尔森(Bozhokina 等人,[2011]Cell Biol Int 35:111-118),从而入侵真核细胞。为了阐明含有格里尔森的细菌入侵如何依赖宿主细胞的生理状态,我们研究了 N-乙酰半胱氨酸 (NAC) 对 HeLa 细胞易感染野生型 S. grimesii 和表达格里尔森基因的重组大肠杆菌的影响。用 10mM NAC 孵育 HeLa 细胞会导致细胞形态的变化和肌动蛋白细胞骨架的解体,当 NAC 从培养基中去除时,这些变化会逆转。在存在 NAC 和去除 NAC 的情况下,产生格里尔森的细菌进入细胞的数量分别增加了 1.5-2 倍和 3-3.5 倍,对于野生型 S. grimesii 和重组大肠杆菌分别如此。这种效应与细胞骨架的重排无关,但可能是由于 NAC 诱导的细胞表面受体上调所致,这些受体在细胞黏附和细胞-细胞连接中发挥作用。S. grimesii 和重组大肠杆菌进入 NAC 处理细胞的效率相差两倍,表明野生型和重组细菌的进入是通过不同的受体进行的,而 NAC 对这些受体的活性有不同的影响。