Mao Xu-Ming, Zhou Zhan, Hou Xiao-Ping, Guan Wen-Jun, Li Yong-Quan
Zhejiang University, College of Life Sciences, Hangzhou 310058, China.
J Bacteriol. 2009 Nov;191(21):6473-81. doi: 10.1128/JB.00875-09. Epub 2009 Sep 4.
Here we reported that deletion of SigK (SCO6520), a sigma factor in Streptomyces coelicolor, caused an earlier switch from vegetative mycelia to aerial mycelia and higher expression of chpE and chpH than that in the wild type. Loss of SigK also resulted in accelerated and enhanced production of antibiotics, actinorhodin, and undecylprodigiosin and increased expression of actII-orf4 and redD. These results suggested that SigK had a negative role in morphological transition and secondary metabolism. Furthermore, the sigK promoter (sigKp) activity gradually increased and sigK expression was partially dependent on SigK, but this dependence decreased during the developmental course of substrate mycelia. Meanwhile, two potentially nonspecific cleavages occurred between SigK and green fluorescent protein, and the SigK fusion proteins expressed under the constitutive promoter ermEp* sharply decreased and disappeared when aerial mycelia emerged. If expressed under sigKp, 3FLAG-SigK showed similar dynamic patterns but did not decrease as sharply as SigK expressed under ermEp*. These data suggested that the climbing expression of sigK might reduce the prompt degradation of SigK during vegetative hypha development for the proper timing of morphogenesis and that SigK vanished to remove the block for the emergence of aerial mycelia. Thus, we proposed that SigK had inhibitory roles on developmental events and that these inhibitory effects may be released by SigK degradation.
在此我们报道,天蓝色链霉菌中的一个σ因子SigK(SCO6520)的缺失导致从营养菌丝体到气生菌丝体的转变提前,且chpE和chpH的表达高于野生型。SigK的缺失还导致抗生素放线紫红素和十一烷基灵菌红素的产生加速且增加,以及actII-orf4和redD的表达增加。这些结果表明SigK在形态转变和次级代谢中起负作用。此外,sigK启动子(sigKp)的活性逐渐增加,sigK的表达部分依赖于SigK,但这种依赖性在底物菌丝体的发育过程中降低。同时,在SigK和绿色荧光蛋白之间发生了两个潜在的非特异性切割,当气生菌丝体出现时,在组成型启动子ermEp下表达的SigK融合蛋白急剧减少并消失。如果在sigKp下表达,3FLAG-SigK显示出类似的动态模式,但不像在ermEp下表达的SigK那样急剧下降。这些数据表明,sigK的逐步表达可能会减少营养菌丝体发育过程中SigK的快速降解,以实现形态发生的适当时间,并且SigK消失以消除气生菌丝体出现的障碍。因此,我们提出SigK对发育事件具有抑制作用,并且这些抑制作用可能通过SigK的降解而解除。