Zhejiang University, Institute of Biochemistry, Hangzhou 310058, China.
Microbiol Res. 2010 Mar 31;165(3):221-31. doi: 10.1016/j.micres.2009.05.003. Epub 2009 Aug 22.
To address the functions of SigN (SCO4034) in Streptomyces coelicolor, we constructed a sigN null mutant M145Z, which showed a defect in sporulation. The differential proteomic profiles of wild-type S. coelicolors M145 and M145Z were demonstrated by two-dimensional polyacrylamide gel electrophoresis (2D PAGE), which identified 24 different spots that were up- or down-regulated due to sigN disruption. Among them, 22 proteins were identified by matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF-MS). The up-regulated proteins were involved in energy metabolism, stress responses, ATP binding, ppGpp stringent stress response and transcriptional anti-termination, etc. The down-regulated proteins were related to antibiotic synthesis and differentiation. The results gave a new insight into the regulatory mechanism of SigN in S. coelicolor. Furthermore, deletion of sigN caused growth retardation under all stress conditions examined including heat, cold, acid, oxidation, salt and ethanol. These results indicated that SigN was involved in morphological development, secondary metabolism and stress responses in S. coelicolor.
为了研究 SigN(SCO4034)在链霉菌中的功能,我们构建了 sigN 缺失突变体 M145Z,该突变体在孢子形成过程中出现缺陷。通过二维聚丙烯酰胺凝胶电泳(2D PAGE)显示了野生型链霉菌 M145 和 M145Z 的差异蛋白质组图谱,由于 sigN 缺失,鉴定出 24 个不同的上调或下调斑点。其中,22 种蛋白质通过基质辅助激光解吸/电离飞行时间质谱(MALDI-TOF-MS)鉴定。上调的蛋白质参与能量代谢、应激反应、ATP 结合、ppGpp 严格应激反应和转录终止等。下调的蛋白质与抗生素合成和分化有关。这些结果为 SigN 在链霉菌中的调控机制提供了新的见解。此外,在所有检测到的应激条件下,包括热、冷、酸、氧化、盐和乙醇,sigN 的缺失都会导致生长迟缓。这些结果表明 SigN 参与了链霉菌的形态发育、次级代谢和应激反应。