Physical Biosciences Division, Lawrence Berkeley National Laboratory Berkeley, CA, USA.
Biochemistry Division, University of Missouri Columbia, MO, USA.
Front Microbiol. 2014 Mar 6;5:77. doi: 10.3389/fmicb.2014.00077. eCollection 2014.
Sulfate-reducing bacteria such as Desulfovibrio vulgaris Hildenborough are often found in environments with limiting growth nutrients. Using lactate as the electron donor and carbon source, and sulfate as the electron acceptor, wild type D. vulgaris shows motility on soft agar plates. We evaluated this phenotype with mutants resulting from insertional inactivation of genes potentially related to motility. Our study revealed that the cheA3 (DVU2072) kinase mutant was impaired in the ability to form motility halos. Insertions in two other cheA loci did not exhibit a loss in this phenotype. The cheA3 mutant was also non-motile in capillary assays. Complementation with a plasmid-borne copy of cheA3 restores wild type phenotypes. The cheA3 mutant displayed a flagellum as observed by electron microscopy, grew normally in liquid medium, and was motile in wet mounts. In the growth conditions used, the D. vulgaris ΔfliA mutant (DVU3229) for FliA, predicted to regulate flagella-related genes including cheA3, was defective both in flagellum formation and in forming the motility halos. In contrast, a deletion of the flp gene (DVU2116) encoding a pilin-related protein was similar to wild type. We conclude that wild type D. vulgaris forms motility halos on solid media that are mediated by flagella-related mechanisms via the CheA3 kinase. The conditions under which the CheA1 (DVU1594) and CheA2 (DVU1960) kinase function remain to be explored.
硫酸盐还原菌,如脱硫弧菌 Hildenborough,通常存在于生长营养物质有限的环境中。利用乳酸作为电子供体和碳源,硫酸盐作为电子受体,野生型 D. vulgaris 在软琼脂平板上表现出运动性。我们用可能与运动性相关的基因插入失活突变体来评估这种表型。我们的研究表明,cheA3 (DVU2072) 激酶突变体在形成运动晕圈的能力上受损。在另外两个 cheA 基因座的插入突变体中,没有表现出这种表型的丧失。cheA3 突变体在毛细管试验中也没有运动性。用质粒携带的 cheA3 拷贝进行互补恢复了野生型表型。cheA3 突变体表现出电子显微镜观察到的鞭毛,在液体培养基中正常生长,在湿载玻片上运动。在使用的生长条件下,用于 FliA 的 D. vulgaris ΔfliA 突变体 (DVU3229),预测调节包括 cheA3 在内的与鞭毛相关基因,在鞭毛形成和运动晕圈形成方面都有缺陷。相比之下,缺失编码 pilin 相关蛋白的 flp 基因 (DVU2116) 与野生型相似。我们得出结论,野生型 D. vulgaris 在固体培养基上形成运动晕圈,这是通过 CheA3 激酶介导的与鞭毛相关的机制。CheA1 (DVU1594) 和 CheA2 (DVU1960) 激酶的作用条件仍有待探索。