Analysis of a taurine-dependent promoter in Sinorhizobium meliloti that offers tight modulation of gene expression

BMC Microbiology

Joseph C Chen¹  Zhenzhong Huang¹  Sean Nicholas King¹  Tuyet Thi Tran¹  Ivan Thomas Gao¹  Julian Albert Bustamante¹  Tanisha Saini¹  Jainee Christa Lewis¹  Mina Mostafavi¹ 

[1] Department of Biology, San Francisco State University, San Francisco 94132, CA, USA

关键词: Twin arginine translocation;    Essential genes;    Taurine metabolism;    mCherry;    Beta-glucuronidase;    Transcriptional regulator;    Sphingomonadaceae;    Rhizobia;    Alphaproteobacteria;   

Others  :  1137723
DOI  :  10.1186/s12866-014-0295-2

 received in 2014-08-27, accepted in 2014-11-13,  发布年份 2014

【 摘 要 】

Background

Genetic models have been developed in divergent branches of the class Alphaproteobacteria to help answer a wide spectrum of questions regarding bacterial physiology. For example, Sinorhizobium meliloti serves as a useful representative for investigating rhizobia-plant symbiosis and nitrogen fixation, Caulobacter crescentus for studying cell cycle regulation and organelle biogenesis, and Zymomonas mobilis for assessing the potentials of metabolic engineering and biofuel production. A tightly regulated promoter that enables titratable expression of a cloned gene in these different models is highly desirable, as it can facilitate observation of phenotypes that would otherwise be obfuscated by leaky expression.

Results

We compared the functionality of four promoter regions in S. meliloti (P_araA, P_tauA, P_rhaR, and P_melA) by constructing strains carrying fusions to the uidA reporter in their genomes and measuring beta-glucuronidase activities when they were induced by arabinose, taurine, rhamnose, or melibiose. P_tauA was chosen for further study because it, and, to a lesser extent, P_melA, exhibited characteristics suitable for efficient modulation of gene expression. The levels of expression from P_tauA depended on the concentrations of taurine, in both complex and defined media, in S. meliloti as well as C. crescentus and Z. mobilis. Moreover, our analysis indicated that TauR, TauC, and TauY are each necessary for taurine catabolism and substantiated their designated roles as a transcriptional activator, the permease component of an ABC transporter, and a major subunit of the taurine dehydrogenase, respectively. Finally, we demonstrated that P_tauA can be used to deplete essential cellular factors in S. meliloti, such as the PleC histidine kinase and TatB, a component of the twin-arginine transport machinery.

Conclusions

The P_tauA promoter of S. meliloti can control gene expression with a relatively inexpensive and permeable inducer, taurine, in diverse alpha-proteobacteria. Regulated expression of the same gene in different hosts can be achieved by placing both tauR and P_tauA on appropriate vectors, thus facilitating inspection of conservation of gene function across species.

【 授权许可】

   
2014 Mostafavi et al.; licensee BioMed Central Ltd.

【 图 表 】

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