Rosu Valentina, Hughes Kelly T
Department of Biology, University of Utah, Salt Lake City, UT 84112, USA.
J Bacteriol. 2006 Jul;188(14):5196-203. doi: 10.1128/JB.00299-06.
The FlgM anti-sigma28 factor is secreted in response to flagellar hook-basal body completion to allow sigma28-dependent transcription of genes needed late in flagellar assembly, such as the flagellin structural gene, fliC. A long-standing hypothesis was that one role of FlgM secretion was to allow rapid expression of flagellin in response to shearing. We tested this hypothesis by following FlgM secretion and fliC transcription in response to flagellar shearing. Experiments showed that the level of FlgM inside the cell was unchanged after shearing whereas the extracellular FlgM levels increased in the growth medium as time passed. Identical results were obtained with cells that were not exposed to shear forces: internal FlgM levels remained constant while external FlgM levels rose with time at rates similar to those for the sheared culture. Consistent with this find, FlgM/sigma28-dependent class 3 gene expression was unaffected by flagellar shearing but was affected by the growth phase of the cell. Regardless of exposure to shear forces, flagellar class 3 transcription rose sharply and then declined. These results demonstrate that flagellar regrowth following shearing is independent of FlgM secretion.
FlgM抗σ28因子在鞭毛钩-基体完成组装时被分泌,从而允许σ28依赖性转录鞭毛组装后期所需的基因,如鞭毛蛋白结构基因fliC。长期以来的一个假说是,FlgM分泌的一个作用是响应剪切力使鞭毛蛋白快速表达。我们通过追踪FlgM分泌和fliC转录对鞭毛剪切的响应来验证这一假说。实验表明,剪切后细胞内FlgM水平未变,而随着时间推移,生长培养基中细胞外FlgM水平升高。未暴露于剪切力的细胞也得到了相同的结果:细胞内FlgM水平保持恒定,而细胞外FlgM水平随时间升高,速率与剪切培养的细胞相似。与此发现一致,FlgM/σ28依赖性3类基因表达不受鞭毛剪切影响,但受细胞生长阶段影响。无论是否暴露于剪切力,鞭毛3类转录均先急剧上升然后下降。这些结果表明,剪切后鞭毛的重新生长与FlgM分泌无关。