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Tracking of single tRNAs for translation kinetics measurements in chloramphenicol treated bacteria
Methods ( IF 4.8 ) Pub Date : 2019-06-01 , DOI: 10.1016/j.ymeth.2019.02.004
Ivan L Volkov 1 , A Carolin Seefeldt 1 , Magnus Johansson 1
Affiliation  

Chloramphenicol is a broad-spectrum antibiotic targeting the protein synthesis machinery by binding to the bacterial ribosome. Chloramphenicol has been considered a classic general inhibitor of translation, blocking the accommodation of aa-tRNA into the A site of the large ribosomal subunit. However, recent studies suggest that this proposed mechanism is a simplification and that the effect of chloramphenicol on mRNA translation is much more dynamic. By tracking single dye-labelled elongator and initiator tRNAs in Escherichia coli cells treated with chloramphenicol, we observe the direct effect of chloramphenicol on translation kinetics. We find clear indications of slow but significant mRNA translation on drug bound ribosomes.

中文翻译:

跟踪单个 tRNA 用于氯霉素处理细菌的翻译动力学测量

氯霉素是一种广谱抗生素,通过与细菌核糖体结合来靶向蛋白质合成机制。氯霉素被认为是一种经典的通用翻译抑制剂,可阻止 aa-tRNA 进入大核糖体亚基的 A 位点。然而,最近的研究表明,这种提议的机制是一种简化,氯霉素对 mRNA 翻译的影响要动态得多。通过在用氯霉素处理的大肠杆菌细胞中跟踪单个染料标记的延伸子和起始 tRNA,我们观察到氯霉素对翻译动力学的直接影响。我们在药物结合的核糖体上发现了缓慢但显着的 mRNA 翻译的明确迹象。
更新日期:2019-06-01
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