Takekawa Norihiro, Kwon Soojin, Nishioka Noriko, Kojima Seiji, Homma Michio
Division of Biological Science, Graduate School of Science, Nagoya University, Chikusa-ku, Nagoya, Japan.
Division of Biological Science, Graduate School of Science, Nagoya University, Chikusa-ku, Nagoya, Japan
J Bacteriol. 2016 Oct 21;198(22):3091-3098. doi: 10.1128/JB.00462-16. Print 2016 Nov 15.
The marine bacterium Vibrio alginolyticus has a single polar flagellum, the number of which is regulated positively by FlhF and negatively by FlhG. FlhF is intrinsically localized at the cell pole, whereas FlhG is localized there through putative interactions with the polar landmark protein HubP. Here we focused on the role of HubP in the regulation of flagellar number in V. alginolyticus Deletion of hubP increased the flagellar number and completely disrupted the polar localization of FlhG. It was thought that the flagellar number is determined primarily by the absolute amount of FlhF localized at the cell pole. Here we found that deletion of hubP increased the flagellar number although it did not increase the polar amount of FlhF. We also found that FlhG overproduction did not reduce the polar localization of FlhF. These results show that the absolute amount of FlhF is not always the determinant of flagellar number. We speculate that cytoplasmic FlhG works as a quantitative regulator, controlling the amount of FlhF localized at the pole, and HubP-anchored polar FlhG works as a qualitative regulator, directly inhibiting the activity of polar FlhF. This regulation by FlhF, FlhG, and HubP might contribute to achieving optimal flagellar biogenesis at the cell pole in V. alginolyticus IMPORTANCE: For regulation of the flagellar number in marine Vibrio, two proteins, FlhF and FlhG, work as positive and negative regulators, respectively. In this study, we found that the polar landmark protein HubP is involved in the regulation of flagellar biogenesis. Deletion of hubP increased the number of flagella without increasing the amount of pole-localizing FlhF, indicating that the number of flagella is not determined solely by the absolute amount of pole-localizing FlhF, which is inconsistent with the previous model. We propose that cytoplasmic FlhG and HubP-anchored polar FlhG negatively regulate flagellar formation through two independent schemes.
溶藻弧菌这种海洋细菌具有一根极鞭毛,其数量受FlhF正向调控,受FlhG负向调控。FlhF本质上定位于细胞极,而FlhG通过与极性地标蛋白HubP的假定相互作用定位于此。在此,我们聚焦于HubP在溶藻弧菌鞭毛数量调控中的作用。hubP缺失增加了鞭毛数量,并完全破坏了FlhG的极性定位。曾认为鞭毛数量主要由定位于细胞极的FlhF的绝对量决定。在此我们发现,hubP缺失增加了鞭毛数量,尽管它并未增加FlhF的极性量。我们还发现,FlhG过量表达并未减少FlhF的极性定位。这些结果表明,FlhF的绝对量并不总是鞭毛数量的决定因素。我们推测,细胞质中的FlhG作为定量调节剂,控制定位于极的FlhF的量,而HubP锚定的极性FlhG作为定性调节剂,直接抑制极性FlhF的活性。FlhF、FlhG和HubP的这种调控可能有助于溶藻弧菌在细胞极实现最佳的鞭毛生物合成。重要性:对于海洋弧菌鞭毛数量的调控,两种蛋白FlhF和FlhG分别作为正向和负向调节剂起作用。在本研究中,我们发现极性地标蛋白HubP参与鞭毛生物合成的调控。hubP缺失增加了鞭毛数量,而未增加定位于极的FlhF的量,这表明鞭毛数量并非仅由定位于极的FlhF的绝对量决定,这与之前的模型不一致。我们提出,细胞质中的FlhG和HubP锚定的极性FlhG通过两种独立机制对鞭毛形成进行负调控。