Quantifying Elongation Rhythm during Full-Length Protein Synthesis

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• 作者：Gabriel Rosenblum ; Chunlai Chen ; Jaskiran Kaur ; Xiaonan Cui ; Haibo Zhang ; Haruichi Asahara ; Shaorong Chong ; Zeev Smilansky ; Yale E. Goldman ; Barry S. Cooperman
• 刊名：The Journal of the American Chemical Society
• 出版年：2013
• 出版时间：July 31, 2013
• 年：2013
• 卷：135
• 期：30
• 页码：11322-11329
• 全文大小：407K
• 年卷期：v.135,no.30(July 31, 2013)
• ISSN：1520-5126

文摘

Pauses regulate the rhythm of ribosomal protein synthesis. Mutations disrupting even minor pauses can give rise to improperly formed proteins and human disease. Such minor pauses are difficult to characterize by ensemble methods, but can be readily examined by single-molecule (sm) approaches. Here we use smFRET to carry out real-time monitoring of the expression of a full-length protein, the green fluorescent protein variant Emerald GFP. We demonstrate significant correlations between measured elongation rates and codon and isoacceptor tRNA usage, and provide a quantitative estimate of the effect on elongation rate of replacing a codon recognizing an abundant tRNA with a synonymous codon cognate to a rarer tRNA. Our results suggest that tRNA selection plays an important general role in modulating the rates and rhythms of protein synthesis, potentially influencing simultaneous co-translational processes such as folding and chemical modification.