Saulmon Michael M, Karatan Ece, Ordal George W
Department of Biochemistry, Colleges of Medicine and Liberal Arts and Sciences, University of Illinois, Urbana, IL 61801, USA.
Microbiology (Reading). 2004 Mar;150(Pt 3):581-589. doi: 10.1099/mic.0.26463-0.
Bacillus subtilis has a more complex mechanism of chemotaxis than does the paradigm organism, Escherichia coli. In order to understand better the role of the novel chemotaxis proteins--CheC, CheD and CheV--mutants in which increasing numbers of the corresponding genes had been deleted were studied as tethered cells and their biases and sometimes durations of counterclockwise (CCW) and clockwise (CW) flagellar rotations in response to addition and removal of the attractant asparagine were observed. The cheC mutant was found to have considerably reduced switching frequency (that is, prolonged CCW and CW rotations) without a significantly different prestimulus CCW bias, compared with wild-type. This result may indicate that in absence of CheC the switch might be in a conformation less resembling the transition state than in presence of CheC. Conversely, the cheB (methylesterase) mutant showed considerably increased switching frequency without affecting CCW bias, compared with wild-type. Removal of all known adaptation systems--the methylation, CheC and CheV systems--resulted in a mutant (cheRBCDV) that still retained some adaptation following the addition of attractant.
与典型生物大肠杆菌相比,枯草芽孢杆菌具有更为复杂的趋化机制。为了更好地理解新型趋化蛋白CheC、CheD和CheV的作用,我们以拴系细胞的形式研究了相应基因缺失数量不断增加的突变体,并观察了它们在添加和去除引诱剂天冬酰胺时逆时针(CCW)和顺时针(CW)鞭毛旋转的偏向以及有时还有持续时间。与野生型相比,发现cheC突变体的转换频率显著降低(即CCW和CW旋转延长),而刺激前的CCW偏向没有显著差异。该结果可能表明,在没有CheC的情况下,开关的构象可能比有CheC时更不像过渡态。相反,与野生型相比,cheB(甲基酯酶)突变体的转换频率显著增加,而不影响CCW偏向。去除所有已知的适应系统——甲基化、CheC和CheV系统——产生了一个突变体(cheRBCDV),该突变体在添加引诱剂后仍保留一些适应性。