Blevins Jon S, Beenken Karen E, Elasri Mohamed O, Hurlburt Barry K, Smeltzer Mark S
Department of Microbiology, University of Arkansas for Medical Sciences, Little Rock, Arkansas 72205, USA.
Infect Immun. 2002 Feb;70(2):470-80. doi: 10.1128/IAI.70.2.470-480.2002.
The accessory gene regulator (agr) and the staphylococcal accessory regulator (sar) are central regulatory elements that control the production of Staphylococcus aureus virulence factors. To date, the functions of these loci have been defined almost exclusively using RN6390, which is representative of the laboratory strain 8325-4. However, RN6390 was recently shown to have a mutation in rsbU that results in a phenotype resembling that of a sigB mutant (I. Kullik et al., J. Bacteriol. 180:4814-4820, 1998). For that reason, it remains unclear whether the regulatory events defined in RN6390 are representative of the events that take place in clinical isolates of S. aureus. To address this issue, we generated mutations in the sarA and agr loci of three laboratory strains (RN6390, Newman, and S6C) and four clinical isolates (UAMS-1, UAMS-601, DB, and SC-1). Mutation of sarA in the cna-positive strains UAMS-1 and UAMS-601 resulted in an increased capacity to bind collagen, while mutation of agr had little impact. Northern blot analysis confirmed that the increase in collagen binding was due to increased cna transcription. Without exception, mutation of sarA resulted in increased production of proteases and a decreased capacity to bind fibronectin. Mutation of agr had the opposite effect. Although mutation of sarA resulted in a slight reduction in fnbA transcription, changes in the ability to bind fibronectin appeared to be more directly correlated with changes in protease activity. Lipase production was reduced in both sarA and agr mutants. While mutation of sarA in RN6390 resulted in reduced hemolytic activity, it had the opposite effect in all other strains. There appeared to be reduced levels of the sarC transcript in RN6390, but there was no difference in the overall pattern of sar transcription or the production of SarA. Although mutation of sarA resulted in decreased RNAIII transcription, this effect was not evident under all growth conditions. Taken together, these results suggest that studies defining the regulatory roles of sarA and agr by using RN6390 are not always representative of the events that occur in clinical isolates of S. aureus.
附属基因调控因子(agr)和葡萄球菌附属调控因子(sar)是控制金黄色葡萄球菌毒力因子产生的核心调控元件。迄今为止,这些基因座的功能几乎完全是通过使用RN6390来定义的,RN6390代表实验室菌株8325-4。然而,最近发现RN6390的rsbU基因发生了突变,导致其表型类似于sigB突变体(I. Kullik等人,《细菌学杂志》180:4814 - 4820,1998年)。因此,尚不清楚在RN6390中定义的调控事件是否代表金黄色葡萄球菌临床分离株中发生的事件。为了解决这个问题,我们在三种实验室菌株(RN6390、Newman和S6C)以及四种临床分离株(UAMS-1、UAMS-601、DB和SC-1)的sarA和agr基因座中产生了突变。在cna阳性菌株UAMS-1和UAMS-601中,sarA突变导致结合胶原蛋白的能力增强,而agr突变影响较小。Northern印迹分析证实,胶原蛋白结合能力的增加是由于cna转录增加所致。无一例外,sarA突变导致蛋白酶产生增加,结合纤连蛋白的能力下降。agr突变则产生相反的效果。虽然sarA突变导致fnbA转录略有减少,但结合纤连蛋白能力的变化似乎与蛋白酶活性的变化更直接相关。在sarA和agr突变体中,脂肪酶的产生均减少。虽然RN6390中的sarA突变导致溶血活性降低,但在所有其他菌株中却产生相反的效果。RN6390中sarC转录本水平似乎降低,但sar转录总体模式或SarA产生没有差异。虽然sarA突变导致RNAIII转录减少,但这种效果在所有生长条件下并不明显。综上所述,这些结果表明,通过使用RN6390来定义sarA和agr调控作用的研究并不总是代表金黄色葡萄球菌临床分离株中发生的事件。