Trip Erik N, Scheffers Dirk-Jan
Department of Molecular Microbiology, Groningen Biomolecular Sciences and Biotechnology Institute, University of Groningen, The Netherlands.
Sci Rep. 2015 Dec 8;5:18190. doi: 10.1038/srep18190.
Cell division in bacteria is an essential process that is carried out at mid-cell by a group of cell division proteins referred to as the divisome. In Escherichia coli, over two dozen cell division proteins have been identified of which ten are essential. These division proteins localize sequentially and interdependently to the division site, after which constriction eventually produces two daughter cells. Various genetic and biochemical techniques have identified many interactions amongst cell division proteins, however the existence of the divisome as a large multi-protein complex has never been shown. Here, we identify a 1 MDa protein complex by native page that contains seven essential cell division proteins (FtsZ, ZipA, FtsK, FtsQ, FtsB, FtsL, and FtsN). The 1 MDa complex is present in rapidly dividing cells, but absent when cultures enter the stationary growth phase. Slight overexpression of the ftsQ D237N mutation that blocks cell division prevents formation of this 1 MDa complex. In cells depleted of FtsN, the 1 MDa complex is not assembled. Combined, our findings indicate that a large protein complex containing many different cell division proteins indeed exists. We note that this complex is very fragile and sensitive to the expression of tagged versions of FtsQ.
细菌中的细胞分裂是一个重要过程,由一组称为分裂体的细胞分裂蛋白在细胞中部进行。在大肠杆菌中,已鉴定出二十多种细胞分裂蛋白,其中十种是必需的。这些分裂蛋白依次且相互依赖地定位于分裂位点,之后缢缩最终产生两个子细胞。各种遗传和生化技术已确定了细胞分裂蛋白之间的许多相互作用,然而,分裂体作为一种大型多蛋白复合物的存在从未得到证实。在这里,我们通过非变性聚丙烯酰胺凝胶电泳鉴定出一种1兆道尔顿的蛋白复合物,它包含七种必需的细胞分裂蛋白(FtsZ、ZipA、FtsK、FtsQ、FtsB、FtsL和FtsN)。这种1兆道尔顿的复合物存在于快速分裂的细胞中,但当培养物进入稳定生长期时则不存在。阻断细胞分裂的ftsQ D237N突变的轻微过表达会阻止这种1兆道尔顿复合物的形成。在缺乏FtsN的细胞中,1兆道尔顿的复合物无法组装。综合来看,我们的研究结果表明确实存在一种包含许多不同细胞分裂蛋白的大型蛋白复合物。我们注意到这种复合物非常脆弱,并且对FtsQ标记版本的表达很敏感。