Jani Charul, Tocheva Elitza I, McAuley Scott, Craney Arryn, Jensen Grant J, Nodwell Justin
Department of Biochemistry and Biomedical Sciences, Michael DeGroote Institute for Infectious Diseases Research, McMaster University, Hamilton, ON, Canada.
Division of Biology and Biological Engineering, California Institute of Technology, Pasadena, CA, USA.
J Biomol Screen. 2015 Feb;20(2):275-84. doi: 10.1177/1087057114551334. Epub 2014 Sep 25.
Cell division is essential for spore formation but not for viability in the filamentous streptomycetes bacteria. Failure to complete cell division instead blocks spore formation, a phenotype that can be visualized by the absence of gray (in Streptomyces coelicolor) and green (in Streptomyces venezuelae) spore-associated pigmentation. Despite the lack of essentiality, the streptomycetes divisome is similar to that of other prokaryotes. Therefore, the chemical inhibitors of sporulation in model streptomycetes may interfere with the cell division in rod-shaped bacteria as well. To test this, we investigated 196 compounds that inhibit sporulation in S. coelicolor. We show that 19 of these compounds cause filamentous growth in Bacillus subtilis, consistent with impaired cell division. One of the compounds is a DNA-damaging agent and inhibits cell division by activating the SOS response. The remaining 18 act independently of known stress responses and may therefore act on the divisome or on divisome positioning and stability. Three of the compounds (Fil-1, Fil-2, and Fil-3) confer distinct cell division defects on B. subtilis. They also block B. subtilis sporulation, which is mechanistically unrelated to the sporulation pathway of streptomycetes but is also dependent on the divisome. We discuss ways in which these differing phenotypes can be used in screens for cell division inhibitors.
细胞分裂对于丝状链霉菌的孢子形成至关重要,但对于其生存能力并非必需。未能完成细胞分裂反而会阻止孢子形成,这种表型可通过缺乏灰色(在天蓝色链霉菌中)和绿色(在委内瑞拉链霉菌中)与孢子相关的色素沉着来观察到。尽管细胞分裂并非必需,但链霉菌的分裂体与其他原核生物的分裂体相似。因此,模型链霉菌中孢子形成的化学抑制剂可能也会干扰杆状细菌的细胞分裂。为了验证这一点,我们研究了196种抑制天蓝色链霉菌孢子形成的化合物。我们发现其中19种化合物会导致枯草芽孢杆菌出现丝状生长,这与细胞分裂受损一致。其中一种化合物是DNA损伤剂,通过激活SOS反应来抑制细胞分裂。其余18种化合物的作用独立于已知的应激反应,因此可能作用于分裂体或分裂体的定位及稳定性。其中三种化合物(Fil-1、Fil-2和Fil-3)会给枯草芽孢杆菌带来不同的细胞分裂缺陷。它们还会阻止枯草芽孢杆菌的孢子形成,这在机制上与链霉菌的孢子形成途径无关,但同样依赖于分裂体。我们讨论了如何利用这些不同的表型来筛选细胞分裂抑制剂。