Department of Pathobiological Sciences, Louisiana State University, Baton Rouge, Louisiana 70803, USA.
J Bacteriol. 2010 Dec;192(23):6105-15. doi: 10.1128/JB.00571-10. Epub 2010 Sep 24.
The gene bb0250 of Borrelia burgdorferi is a homolog of the dedA family, encoding integral inner membrane proteins that are present in nearly all species of bacteria. To date, no precise function has been attributed to any dedA gene. Unlike many bacterial species, such as Escherichia coli, which has eight dedA genes, B. burgdorferi possesses only one, annotated bb0250, providing a unique opportunity to investigate the functions of the dedA family. Here, we show that bb0250 is able to restore normal growth and cell division to a temperature-sensitive E. coli mutant with simultaneous deletions of two dedA genes, yqjA and yghB, and encodes a protein that localizes to the inner membrane of E. coli. The bb0250 gene could be deleted from B. burgdorferi only after introduction of a promoterless bb0250 under the control of an inducible lac promoter, indicating that it is an essential gene in this organism. Growth of the mutant in the absence of isopropyl-β-d-thiogalactopyranoside resulted in cell death, preceded by cell division defects characterized by elongated cells and membrane bulges, demonstrating that bb0250 is required for proper cell division and envelope integrity. Finally, we show that BB0250 depletion leads to imbalanced membrane phospholipid composition in borrelia. These results demonstrate a strong conservation of function of the dedA gene family across diverse species of Gram-negative bacteria and a requirement for this protein family for normal membrane lipid composition and cell division.
伯氏疏螺旋体的 bb0250 基因是 dedA 家族的同源物,编码存在于几乎所有细菌物种中的完整内膜蛋白。迄今为止,尚未赋予任何 dedA 基因确切的功能。与许多细菌物种(如具有 8 个 dedA 基因的大肠杆菌)不同,伯氏疏螺旋体仅拥有一个,注释为 bb0250,这为研究 dedA 家族的功能提供了独特的机会。在这里,我们表明 bb0250 能够恢复同时缺失两个 dedA 基因(yjjA 和 yghB)的温度敏感型大肠杆菌突变体的正常生长和细胞分裂,并编码一种定位于大肠杆菌内膜的蛋白质。只有在引入无启动子的 bb0250 基因并受诱导型 lac 启动子控制后,才能从伯氏疏螺旋体中删除 bb0250 基因,表明它是该生物体内的必需基因。在没有异丙基-β-D-硫代半乳糖吡喃糖苷的情况下,突变体的生长导致细胞死亡,这之前的特征是细胞分裂缺陷,表现为细胞伸长和膜膨出,表明 bb0250 是正常细胞分裂和包膜完整性所必需的。最后,我们表明 BB0250 耗竭会导致伯氏疏螺旋体中膜磷脂组成失衡。这些结果表明,dedA 基因家族在不同的革兰氏阴性细菌物种中具有很强的功能保守性,并且该蛋白家族对于正常的膜脂组成和细胞分裂是必需的。