Schirner Kathrin, Errington Jeff
Institute for Cell and Molecular Biosciences, Newcastle University, Newcastle Upon Tyne, United Kingdom.
J Bacteriol. 2009 Mar;191(5):1404-13. doi: 10.1128/JB.01497-08. Epub 2008 Dec 29.
Bacterial actin homologues are thought to have a role in cell shape determination by positioning the cell wall synthetic machinery. They are also thought to control other functions, including cell polarity and chromosome segregation in various organisms. Bacillus subtilis and many other gram-positive bacteria have three actin isoforms, MreB, Mbl, and MreBH, which colocalize in helical structures that span the length of the cell, close to the inner surface of the cytoplasmic membrane. Deletion of the mbl gene has previously been reported to produce viable, although poorly growing, mutant cells. We now show that under normal conditions Deltambl cells are nonviable but suppressors allowing growth readily accumulate. In the presence of high concentrations of Mg(2+), viable, nonsuppressed mutants can be obtained. A screen for suppressor mutations revealed that deletion of rsgI restores Mg(2+)-independent growth of the mbl mutant. Recent work has shown that rsgI deletion leads to upregulation of the alternative sigma factor sigma(I). The basis of suppression is not yet clear, but it is independent of the Mg(2+) effect. We found that the construction of a triple mutant lacking all three actin homologues became possible in the rsgI background. Triple mutant cells are spherical, but no significant defect in chromosome segregation was detected.
细菌肌动蛋白同源物被认为通过定位细胞壁合成机制在细胞形状决定中发挥作用。它们还被认为控制其他功能,包括各种生物体中的细胞极性和染色体分离。枯草芽孢杆菌和许多其他革兰氏阳性细菌有三种肌动蛋白异构体,MreB、Mbl和MreBH,它们共定位于跨越细胞长度的螺旋结构中,靠近细胞质膜的内表面。先前有报道称,mbl基因的缺失会产生存活但生长不良的突变细胞。我们现在表明,在正常条件下,Δmbl细胞无法存活,但允许生长的抑制子很容易积累。在高浓度Mg(2+)存在的情况下,可以获得存活的、未被抑制的突变体。对抑制子突变的筛选表明,rsgI的缺失恢复了mbl突变体不依赖Mg(2+)的生长。最近的研究表明,rsgI的缺失导致替代西格玛因子西格玛(I)的上调。抑制的基础尚不清楚,但它与Mg(2+)效应无关。我们发现在rsgI背景下构建缺乏所有三种肌动蛋白同源物的三重突变体成为可能。三重突变体细胞呈球形,但未检测到染色体分离的明显缺陷。