Koroyasu S, Yamazato M, Hirano T, Aizawa S I
Department of Mechanical and Precision Systems, Teikyo University, Utsunomiya, Japan.
Biophys J. 1998 Jan;74(1):436-43. doi: 10.1016/S0006-3495(98)77801-4.
The growth rate of flagellar hooks in Salmonella typhimurium was analyzed by computer-aided simulation of the length distributions of mutant hooks of uncontrolled length (polyhooks). The wild-type hook has a relatively well-controlled length, with an average of 55 nm and a standard deviation of 6 nm. Mutations in the fliK gene give rise to polyhooks. A histogram of the lengths of polyhooks from a fliK mutant shows a peak at 55 nm with a long monotonic tail extending out to 1 microm. To analyze the growth rate, we employed the population balance method. Regression analysis showed that the histogram could fit a combination of two theoretical curves. In the first phase of growth, the hook starts with a very fast growth rate (40 nm/min), and then the rate exponentially slows until the length reaches 55 nm. In the second phase of growth, where the hook length is over 55 nm, the hook grows at a constant rate of 8 nm/min. Second mutations in either the fliK or flhB genes, as found in pseudorevertants from fliK mutants, give rise to polyhook filaments (phf). The ratio between the numbers of hooks with and without filament was 6:4. The calculated probability of filament attachment to polyhooks was low so that the proportion of hooks that start filament growth was only 2% per minute. The lengths of polyhooks with and without filaments were measured. A histogram of hook length in phf's was the same as that for polyhooks in single-site fliK mutants, against the expectation that the distribution would shift to a shorter average. The role of FliK in hook length control is discussed.
通过对长度不受控制的突变型鞭毛钩(多钩)长度分布进行计算机辅助模拟,分析了鼠伤寒沙门氏菌中鞭毛钩的生长速率。野生型鞭毛钩长度控制相对良好,平均长度为55纳米,标准差为6纳米。fliK基因突变会产生多钩。来自fliK突变体的多钩长度直方图在55纳米处有一个峰值,并有一条长长的单调尾巴延伸至1微米。为了分析生长速率,我们采用了群体平衡法。回归分析表明,该直方图可以拟合两条理论曲线的组合。在生长的第一阶段,鞭毛钩开始时生长速率非常快(40纳米/分钟),然后速率呈指数级减慢,直到长度达到55纳米。在生长的第二阶段,当鞭毛钩长度超过55纳米时,鞭毛钩以8纳米/分钟的恒定速率生长。在fliK突变体的假回复体中发现,fliK或flhB基因的第二次突变会产生多钩丝(phf)。有丝和无丝钩的数量之比为6:4。计算得出丝附着在多钩上的概率较低,因此开始丝生长的钩的比例仅为每分钟2%。测量了有丝和无丝多钩的长度。phf中钩长度的直方图与单一位点 fliK突变体中多钩的直方图相同,这与预期的分布会向较短平均值偏移的情况相反。讨论了FliK在钩长度控制中的作用。