Ranhand J M
J Bacteriol. 1974 Jun;118(3):1041-50. doi: 10.1128/jb.118.3.1041-1050.1974.
Reagents that interact with sulfhydryl groups are shown to inhibit competence factor (CF)-induced competence development in Streptococcus sanguis (Wicky) strain WE4 (Wicky 4 Ery(R)). Inhibition is correlated with specific inhibition of either the function or biosynthesis of three competent cell-related proteins and is reversed by either 2-mercaptoethanol or dithiothreitol. Mercuric chloride (5 muM) or N-ethylmaleimide (NEM; 50 muM) inhibited (i) the function but not the biosynthesis or activation of the competent cell-associated autolysin; (ii) the biosynthesis of a competent cell-associated protein of unknown function, demonstrated by polyacrylamide gel electrophoresis of acidified phenol extracts; and (iii) the biosynthesis or activation of distinct deoxyribonucleic acid (DNA)-binding sites. Neither reagent at the indicated concentration interfered with the uptake of CF by cells or with the uptake and expression of DNA by competent cells. Neither reagent inactivated CF or genetic markers coded by the transforming DNA, nor did they inhibit cell growth or viability appreciably. The data reveal that either mercuric chloride or NEM can differentially inhibit induced protein synthesis and, in addition, conclusively show that some autolytic activity is essential for the onset of the competent state.
已表明与巯基相互作用的试剂会抑制血链球菌(威基)菌株WE4(威基4 红霉素抗性)中感受态因子(CF)诱导的感受态发育。抑制作用与三种感受态细胞相关蛋白的功能或生物合成的特异性抑制相关,并且可被2-巯基乙醇或二硫苏糖醇逆转。氯化汞(5 μM)或N-乙基马来酰亚胺(NEM;50 μM)抑制:(i)感受态细胞相关自溶素的功能,但不抑制其生物合成或激活;(ii)一种功能未知的感受态细胞相关蛋白的生物合成,这通过酸化苯酚提取物的聚丙烯酰胺凝胶电泳得以证明;以及(iii)不同的脱氧核糖核酸(DNA)结合位点的生物合成或激活。在所示浓度下,这两种试剂均不干扰细胞对CF的摄取或感受态细胞对DNA的摄取及表达。这两种试剂均未使CF或由转化DNA编码的遗传标记失活,它们也未明显抑制细胞生长或活力。数据表明,氯化汞或NEM均可差异性地抑制诱导的蛋白质合成,此外,确凿地表明某些自溶活性对于感受态状态的起始至关重要。