Genome-wide analysis in vivo of translation with nucleotide resolution using ribosome profiling.
Genome-wide analysis in vivo of translation with nucleotide resolution using ribosome profiling.
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全基因组分析在体内翻译,并使用核糖体谱分辨率分辨出核苷酸分析。
DOI:
10.1126/science.1168978
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发表时间:
2009-04-10
期刊:
影响因子:
--
通讯作者:
Weissman JS
中科院分区:
文献类型:
--
作者:
Ingolia NT;Ghaemmaghami S;Newman JR;Weissman JS
Techniques for systematically monitoring protein translation have lagged far behind methods for measuring messenger RNA (mRNA) levels. Here, we present a ribosome-profiling strategy that is based on the deep sequencing of ribosome-protected mRNA fragments and enables genome-wide investigation of translation with subcodon resolution. We used this technique to monitor translation in budding yeast under both rich and starvation conditions. These studies defined the protein sequences being translated and found extensive translational control in both determining absolute protein abundance and responding to environmental stress. We also observed distinct phases during translation that involve a large decrease in ribosome density going from early to late peptide elongation as well as widespread regulated initiation at non—adenine-uracil-guanine (AUG) codons. Ribosome profiling is readily adaptable to other organisms, making high-precision investigation of protein translation experimentally accessible.
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