He Xiangyun, Thornton Justin, Carmicle-Davis Stephanie, McDaniel Larry S
Department of Microbiology, University of Mississippi Medical Center, Jackson, 2500 North State Street, MS 39216, USA.
Microb Pathog. 2006 Dec;41(6):199-206. doi: 10.1016/j.micpath.2006.07.001. Epub 2006 Sep 25.
We have identified a pneumococcal gene, tex, which has the potential to regulate gene expression. The tex gene is named for its role in toxin expression in Bordetella pertussis, where it was characterized as an essential gene. Homologous sequences have been found in both Gram-positive and Gram-negative bacteria and are highly conserved at the protein level. Tex family proteins contain a S1 RNA-binding domain at the C-terminus. Members of this family are putative transcriptional accessory factors. Although tex in Streptococcus pneumoniae is homologous to that in B. pertussis, there are distinct differences. Since the tex gene in S. pneumoniae is not an essential gene, we were able to delete tex in strain D39. The tex knockout mutant, DeltaTex, did not affect production of the pneumococcal toxin pneumolysin. However, we observed decreased growth of DeltaTex in the presence of the wild-type strain both in vitro and in vivo as determined by generation numbers and competitive index (CI). The interaction between recombinant Tex and nucleic acids was confirmed by southwestern and northwestern analysis, supporting its role as a transcriptional accessory factor.
我们已经鉴定出一种肺炎球菌基因tex,它具有调节基因表达的潜力。tex基因因其在百日咳博德特氏菌毒素表达中的作用而得名,在那里它被鉴定为一个必需基因。在革兰氏阳性菌和革兰氏阴性菌中均发现了同源序列,并且在蛋白质水平上高度保守。Tex家族蛋白在C端含有一个S1 RNA结合结构域。该家族成员是假定的转录辅助因子。尽管肺炎链球菌中的tex与百日咳博德特氏菌中的tex同源,但仍存在明显差异。由于肺炎链球菌中的tex基因不是必需基因,我们能够在D39菌株中删除tex。tex基因敲除突变体DeltaTex不影响肺炎球菌毒素肺炎溶血素的产生。然而,通过世代数和竞争指数(CI)测定,我们观察到在体外和体内,野生型菌株存在时DeltaTex的生长均有所下降。通过蛋白质免疫印迹和RNA免疫沉淀分析证实了重组Tex与核酸之间的相互作用,支持了其作为转录辅助因子的作用。