非编码小RNA BSR0602在布鲁菌环境适应能力中的调控作用  

The Role of Small Non-coding RNA BSR0602 in the Regulation of Stress Response in Brucella melitensis

作　　者：袁久云[1,2] 龚春丽[2] 庄妤冰 雷霜霜 陈泽良[2] 黄克和[1] 王玉飞[3,2] 

机构地区：[1]南京农业大学动物医学院,南京210095 [2]军事医学科学院疾病预防控制所,北京100071 [3]武警总医院检验科,北京100039

出　　处：《中国生物化学与分子生物学报》2015年第9期989-998,共10页Chinese Journal of Biochemistry and Molecular Biology

基　　金：国家自然科学基金(No.31000041,No.81171530);北京市科技新星项目(No.Z131102000413062)~~

摘　　要：BSR0602是位于布鲁菌染色体上的非编码小RNA,在前期研究中我们发现,BSR0602与布鲁菌的胞内生存能力相关.为了进一步研究BSR0602对布鲁菌胞内环境适应能力的调控作用,采用双向电泳技术对布鲁菌野生株16M和BSR0602过表达株的全菌蛋白质谱进行比较分析.结果显示,BSR0602过表达后,布鲁菌转运代谢蛋白和压力适应蛋白的表达发生变化.qRT-PCR和HIS表位标记实验结果进一步证实,BSR0602在转录和翻译水平均影响氧压力适应蛋白Sod A的表达.相关表型实验结果显示,BSR0602过表达株对氧压力更为敏感,证实了BSR0602在布鲁菌适应氧压力中的作用.结果表明,非编码小RNA BSR0602作为布鲁菌的转录后调控因子,可通过调控压力适应蛋白的表达来影响布鲁菌的压力适应能力和胞内生存.BSR0602 is a small non-coding RNA located in chromosome II of Brucella melitensis,which is proven to be associated with Brucella intracellular survival. To further study the role of BSR0602 in the regulation of stress response and determine genes regulated by this sRNA,the proteomes of wild-type and BSR0602 overexpression mutant were compared by using two-dimensional polyacrylamide gel electrophoresis（ 2D-PAGE）. The differentially expressed proteins were identified by MALDI-TOF-MS.Most of these proteins participate in amino acid, inorganic ion, and carbohydrate transport and metabolism. Of note is that stress response protein Htr A（ BMEI0613） and SodA（ BMEI1367） were downregulated when BSR0602 was overexpressed. Results of qRT-PCR and Western blot confirmed that BSR0602 inhibition the transcription of sod A. Further phenotype assay demonstrated that BSR0602 overexpression strain were sensitive to endogenous superoxide and H2O2. As a post-transcriptional regulator, small non-coding RNA BSR0602 may play a role in Brucella intracellular survival andenvironment stress adaption by modulating expression of stress response proteins.

关 键 词：布鲁菌 非编码小RNA 双向凝胶电泳 转录调控 氧压力 

分 类 号：R34[医药卫生—基础医学]