Abstract
During translation, intracellular mRNA folds co-transcriptionally and must refold following the passage of ribosome. The mRNAs can be entrapped in metastable structures during these folding events. In the present study, we evaluated the conformational dynamics of the kinetically favored, metastable, and hairpin-like structure, which disturbs the thermodynamically favored G-quadruplex structure, and its effect on co-transcriptional translation in prokaryotic cells. We found that nascent mRNA forms a metastable hairpin-like structure during co-transcriptional folding instead of the G-quadruplex structure. When the translation progressed co-transcriptionally before the metastable hairpin-like structure transition to the G-quadruplex, function of the G-quadruplex as a roadblock of the ribosome was sequestered. This suggested that kinetically formed RNA structures had a dominant effect on gene expression in prokaryotes. The results of this study indicate that it is critical to consider the conformational dynamics of RNA-folding to understand the contributions of the mRNA structures in controlling gene expression.
Highlights
An mRNA sequence determines the sequence of amino acids in protein and controls gene expression by the formation of secondary and tertiary structures [1,2,3,4]
We demonstrated that the metastable hairpin-like structure formed co-transcriptionally or co-translationally and the time lag between transcription and translation are the key factors that affect formation of the G-quadruplex, and thereby, suppression of translation both in vitro and in E. coli cells
We expected a dynamic behavior of the G-rich sequence during its involvement in the processes of gene expression in the cells
Summary
An mRNA sequence determines the sequence of amino acids in protein and controls gene expression by the formation of secondary and tertiary structures [1,2,3,4]. Owing to the directional nature of the transcription reaction, RNA structure elements are folded from 50 to 30 direction, a process referred to as co-transcriptional folding [9,10]. Co-transcriptional RNA folding restricts the landscape of its structure formation, which engages the formation of metastable. These metastable structures are transient and show conformational transition to thermodynamically stable ones during the directional folding or post-transcriptional organization of the RNAs. The structures of coding regions of mRNAs are reorganized co-translationally following passage of the ribosome, which incorporates and discharges single-stranded mRNA as the template strand. The intracellular RNA structures are transient and fluctuate dynamically over time as the processes involved in gene expressions occur
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