Moreno S, Ertesvåg H, Valla S, Núñez C, Espin G, Cocotl-Yañez M
Departamento de Microbiología Molecular, Instituto de Biotecnología, Universidad Nacional Autónoma de México. Av. Universidad 2001, Col. Chamilpa, Cuernavaca, Morelos. C.P. 62210, México.
Department of Biotechnology and Food Science, NTNU-Norwegian University of Science and Technology, Sem Sælands vei 6/8, 7491 Trondheim, Norway.
FEMS Microbiol Lett. 2018 Oct 1;365(19). doi: 10.1093/femsle/fny210.
Azotobacter vinelandii produces differentiated cells, called cysts, surrounded by two alginate layers, which are necessary for their desiccation resistance. This alginate contains variable proportions of guluronate residues, resulting from the activity of seven extracytoplasmic epimerases, AlgE1-7. These enzymes are exported by a system secretion encoded by the eexDEF operon; mutants lacking the AlgE1-7 epimerases, the EexDEF or the RpoS sigma factor produce alginate, but are unable to form desiccation resistant cysts. Herein, we found that RpoS was required for full transcription of the algE1-7 and eexDEF genes. We found that the AlgE1-7 protein levels were diminished in the rpoS mutant strain. In addition, the alginate produced in the absence of RpoS was more viscous in the presence of proteases, a phenotype similar to that of the eexD mutant. Primer extension analysis located two promoters for the eexDEF operon, one of them was RpoS-dependent. Thus, during encysting conditions, RpoS coordinates the expression of both the AlgE1-7 epimerases and the EexDEF protein complex responsible for their transport.
棕色固氮菌会产生一种分化细胞,称为孢囊,其被两层藻酸盐包围,这两层藻酸盐对于孢囊的抗干燥能力至关重要。这种藻酸盐含有不同比例的古洛糖醛酸残基,这是由七种胞外差向异构酶AlgE1 - 7的活性所导致的。这些酶通过由eexDEF操纵子编码的系统分泌途径输出;缺乏AlgE1 - 7差向异构酶、EexDEF或RpoS σ因子的突变体能够产生藻酸盐,但无法形成抗干燥的孢囊。在此,我们发现RpoS是algE1 - 7和eexDEF基因完全转录所必需的。我们发现RpoS突变菌株中AlgE1 - 7蛋白水平降低。此外,在没有RpoS的情况下产生的藻酸盐在蛋白酶存在时更具粘性,这一表型与eexD突变体相似。引物延伸分析确定了eexDEF操纵子的两个启动子,其中一个依赖于RpoS。因此,在形成孢囊的条件下,RpoS协调AlgE1 - 7差向异构酶和负责其转运的EexDEF蛋白复合物的表达。