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FlbT是新月柄杆菌中鞭毛蛋白合成的转录后调节因子,它与鞭毛蛋白mRNA的5'非翻译区相互作用。

FlbT, the post-transcriptional regulator of flagellin synthesis in Caulobacter crescentus, interacts with the 5' untranslated region of flagellin mRNA.

作者信息

Anderson P E, Gober J W

机构信息

Department of Microbiology and Molecular Genetics, University of California, Los Angeles, CA 90095-1569, USA.

出版信息

Mol Microbiol. 2000 Oct;38(1):41-52. doi: 10.1046/j.1365-2958.2000.02108.x.

Abstract

Flagellar gene expression' in Caulobacter crescentus is regulated by a complex trans-acting hierarchy, in which the assembly of early structural proteins is required for the expression of later structural proteins. The flagellins that comprise the filament are regulated at both the transcriptional and the post-transcriptional levels. Post-transcriptional regulation is sensitive to the assembly of the flagellar basal body and hook structures. In mutant strains lacking these structures, flagellin genes are transcribed, but not translated. Mutations in the flagellar regulatory gene, flbT, restore flagellin translation in the absence of flagellar assembly. In this report, we investigate the mechanism of FlbT-mediated post-transcriptional regulation. We show that FlbT is associated with the 5' untranslated region (UTR) of fljK (25 kDa flagellin) mRNA and that this association requires a predicted loop structure in the transcript. Mutations within this loop abolished FlbT association and resulted in increased mRNA stability, indicating that FlbT promotes the degradation of flagellin mRNA by associating with the 5' UTR. We also assayed the effects on gene expression using mutant transcripts fused to lacZ. Interestingly, the mutant transcript that failed to associate with FlbT in vitro was still repressed in mutants defective in flagellum assembly, suggesting that other factors in addition to FlbT couple assembly to translation.

摘要

新月柄杆菌中鞭毛基因的表达受一个复杂的反式作用层级调控,其中早期结构蛋白的组装是后期结构蛋白表达所必需的。构成鞭毛丝的鞭毛蛋白在转录和转录后水平均受到调控。转录后调控对鞭毛基体和钩状结构的组装敏感。在缺乏这些结构的突变菌株中,鞭毛蛋白基因可以转录,但不能翻译。鞭毛调控基因flbT的突变可在缺乏鞭毛组装的情况下恢复鞭毛蛋白的翻译。在本报告中,我们研究了FlbT介导的转录后调控机制。我们发现FlbT与fljK(25 kDa鞭毛蛋白)mRNA的5'非翻译区(UTR)相关联,并且这种关联需要转录本中一个预测的环结构。该环内的突变消除了FlbT的关联,并导致mRNA稳定性增加,这表明FlbT通过与5'UTR结合促进鞭毛蛋白mRNA的降解。我们还使用与lacZ融合的突变转录本检测了对基因表达的影响。有趣的是,在体外未能与FlbT结合的突变转录本在鞭毛组装缺陷的突变体中仍然受到抑制,这表明除了FlbT之外,还有其他因素将组装与翻译联系起来。

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