The State Key Laboratory of Freshwater Ecology and Biotechnology, Institute of Hydrobiology, Chinese Academy of Sciences, 430072, Wuhan, Hubei, China.
Arch Microbiol. 2010 Jun;192(6):461-9. doi: 10.1007/s00203-010-0573-9. Epub 2010 Apr 20.
A protein of cyAbrB family of regulators, Alr0946 (CalA), was identified by DNA affinity chromatography using DNA fragments from the promoter region of ftsZ in Anabaena/Nostoc sp. PCC 7120. Electrophoretic mobility shift assays showed that the protein could bind to upstream regions of ftsZ (as biotin-labeled DNA fragments) and that the binding was inhibited by the same DNA fragments and a fragment upstream of hetC but not by 3 fragments upstream of glnA, hetR or ntcA. Both transcriptional fusion with gfp (green fluorescence protein) and Western blot analysis indicated that the expression of calA was down-regulated in heterocysts relative to vegetative cells. An insertional mutant of calA could not be completely segregated. Over-expression of calA in Anabaena/Nostoc caused no change in growth, heterocyst differentiation or expression of ftsZ. CalA as a DNA-binding protein is essential for the growth of Anabaena/Nostoc and may be required for the expression of ftsZ in vegetative cells.
一种 CyAbrB 家族调控蛋白 Alr0946(CalA),是通过使用 Anabaena/Nostoc sp. PCC 7120 的 ftsZ 启动子区域的 DNA 片段进行 DNA 亲和层析鉴定的。电泳迁移率变动分析表明,该蛋白可以与 ftsZ 的上游区域(如生物素标记的 DNA 片段)结合,并且这种结合可以被相同的 DNA 片段和 hetC 上游的片段抑制,但不能被 glnA、hetR 或 ntcA 上游的 3 个片段抑制。与 gfp(绿色荧光蛋白)的转录融合和 Western blot 分析表明,与营养细胞相比,calA 的表达在异形胞中受到下调。calA 的插入突变体不能完全分离。在 Anabaena/Nostoc 中过表达 calA 不会导致生长、异形胞分化或 ftsZ 的表达发生变化。作为一种 DNA 结合蛋白的 CalA 对 Anabaena/Nostoc 的生长是必不可少的,并且可能是营养细胞中 ftsZ 表达所必需的。