ADHESION PROPERTIES OF MUTANTS OF STAPHYLOCOCCUS-AUREUS DEFECTIVE IN FIBRONECTIN-BINDING PROTEINS AND STUDIES ON THE EXPRESSION OF FNB GENES
ADHESION PROPERTIES OF MUTANTS OF STAPHYLOCOCCUS-AUREUS DEFECTIVE IN FIBRONECTIN-BINDING PROTEINS AND STUDIES ON THE EXPRESSION OF FNB GENES
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DOI：
10.1111/j.1365-2958.1995.mmi_17061143.x
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发表时间：
1995-09-01
影响因子：
3.6
通讯作者：
FOSTER, TJ
中科院分区：
文献类型：
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作者：
GREENE, C;MCDEVITT, D;FOSTER, TJ
金黄色葡萄球菌 8325-4 有可能表达两种不同的细胞壁相关纤连蛋白结合蛋白，称为 FnBPA 和 FnBPB。为了测试这两种蛋白是否在金黄色葡萄球菌中表达，以及这两种蛋白是否都是促进细菌粘附到纤连蛋白包被表面所必需的，在每个基因中分离了插入突变。将编码四环素抗性的DNA片段插入fnbA中，并将编码红霉素抗性的片段插入fnbB中。还构建了双 fnbA fnbB 突变体。 fnbA 和 fnbB 单突变体对体外涂有纤连蛋白的聚甲基丙烯酸甲酯盖玻片的粘附力没有显着降低。然而，双突变体在粘附方面完全有缺陷。针对两种蛋白非保守 N 末端区域的单特异性抗体证实突变菌株中缺乏 FnBP 表达。单独克隆在多拷贝质粒上的野生型fnbA和fnbB基因各自能够完全恢复8325-4 fnbA fnbB突变体的粘附缺陷表型。这表明，两种 fnb 基因均在金黄色葡萄球菌中表达，并且都有助于菌株 8325-4 粘附于纤连蛋白包被表面的能力。双突变体在与盖玻片的粘附方面也存在缺陷，所述盖玻片已从皮下植入豚鼠的组织笼中取出，这表明纤连蛋白对于促进金黄色葡萄球菌粘附至纤连蛋白具有重要作用。 体内生物材料。
Staphylococcus aureus 8325-4 has the potential to express two distinct cell wall-associated fibronectin-binding proteins called FnBPA and FnBPB. In order to test if both proteins are expressed in S. aureus and if both are required for promoting bacterial adhesion to fibronectin-coated surfaces, insertion mutations were isolated in each gene. A DNA fragment encoding tetracycline resistance was inserted into fnbA and a fragment encoding erythromycin resistance was inserted into fnbB. A double fnbA fnbB mutant was also constructed. The fnbA and fnbB single mutants showed no significant reduction in their adhesion to polymethylmethacrylate coverslips that had been coated in vitro with fibronectin. However, the double mutant was completely defective in adhesion. Monospecific antibodies directed against the non-conserved N-terminal regions of both proteins confirmed the lack of expression of FnBPs in the mutant strains. Wild-type fnbA and fnbB genes cloned seperately on a multicopy plasmid were each able to restore fully the adhesion-defective phenotype of the 8325-4 fnbA fnbB mutant. This demonstrates that both fnb genes are expressed in S. aureus and that both contribute to the ability of strain 8325-4 to adhere to fibronectin-coated surfaces, The double mutant was also defective in adhesion to coverslips that had been removed from tissue cages implanted subcutaneously in guinea-pigs, which suggests that fibronectin is important in promoting attachment of S. aureus to biomaterial in vivo.