Wang Sheng-Lan, Fan Ke-Qiang, Yang Xu, Lin Zeng-Xi, Xu Xin-Ping, Yang Ke-Qian
State Key Laboratory of Microbial Resources, Institute of Microbiology, Chinese Academy of Sciences, Beijing 100101, People's Republic of China.
J Bacteriol. 2008 Jun;190(11):4061-8. doi: 10.1128/JB.01954-07. Epub 2008 Mar 28.
Ca(2+) was reported to regulate spore germination and aerial hypha formation in streptomycetes; the underlying mechanism of this regulation is not known. cabC, a gene encoding an EF-hand calcium-binding protein, was disrupted or overexpressed in Streptomyces coelicolor M145. On R5- agar, the disruption of cabC resulted in denser aerial hyphae with more short branches, swollen hyphal tips, and early-germinating spores on the spore chain, while cabC overexpression significantly delayed development. Manipulation of the Ca(2+) concentration in R5- agar could reverse the phenotypes of cabC disruption or overexpression mutants and mimic mutant phenotypes with M145, suggesting that the mutant phenotypes were due to changes in the intracellular Ca(2+) concentration. CabC expression was strongly activated in aerial hyphae, as determined by Western blotting against CabC and confocal laser scanning microscopy detection of CabC::enhanced green fluorescent protein (EGFP). CabC::EGFP fusion proteins were evenly distributed in substrate mycelia, aerial mycelia, and spores. Taken together, these results demonstrate that CabC is involved in Ca(2+)-mediated regulation of spore germination and aerial hypha formation in S. coelicolor. CabC most likely acts as a Ca(2+) buffer and exerts its regulatory effects by controlling the intracellular Ca(2+) concentration.
据报道,Ca(2+)可调节链霉菌的孢子萌发和气生菌丝形成;但这种调节的潜在机制尚不清楚。cabC是一个编码EF-手型钙结合蛋白的基因,在天蓝色链霉菌M145中被破坏或过表达。在R5-琼脂培养基上,cabC的破坏导致气生菌丝更密集,有更多短分支、肿胀的菌丝尖端以及孢子链上的孢子提前萌发,而cabC的过表达则显著延迟了发育。改变R5-琼脂培养基中的Ca(2+)浓度可以逆转cabC破坏或过表达突变体的表型,并模拟M145的突变体表型,这表明突变体表型是由于细胞内Ca(2+)浓度的变化所致。通过针对CabC的蛋白质免疫印迹和共聚焦激光扫描显微镜检测CabC::增强型绿色荧光蛋白(EGFP),发现CabC在气生菌丝中强烈激活。CabC::EGFP融合蛋白均匀分布在基内菌丝、气生菌丝和孢子中。综上所述,这些结果表明CabC参与了Ca(2+)介导的天蓝色链霉菌孢子萌发和气生菌丝形成调控。CabC很可能作为一种Ca(2+)缓冲剂,通过控制细胞内Ca(2+)浓度发挥其调节作用。