글번호
5550043
일 자
16.07.08
조회수
67
글쓴이
김은아
제목 : 2016-07-01 Real-time quantification of single RNA translation dynamics in living cells. (김은아)
2016 Jun 17;352(6292):1425-9. doi: 10.1126/science.aaf0899. Epub 2016 May 5.
 

Real-time quantification of single RNA translation dynamics in living cells.

Abstract

Although messenger RNA (mRNA) translation is a fundamental biological process, it has never been imaged in real time in vivo with single-molecule precision. To achieve this, we developed nascent chain tracking (NCT), a technique that uses multi-epitope tags and antibody-based fluorescent probes to quantify protein synthesis dynamics at the single-mRNA level. NCT reveals an elongation rate of ~10 amino acids per second, with initiation occurring stochastically every ~30 seconds. Polysomes contain ~1 ribosome every 200 to 900 nucleotides and are globular rather than elongated in shape. By developing multicolor probes, we showed that most polysomes act independently; however, a small fraction (~5%) form complexes in which two distinct mRNAs can be translated simultaneously. The sensitivity and versatility of NCT make it a powerful new tool for quantifying mRNA translation kinetics.


Copyright © 2016, American Association for the Advancement of Science.

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