A pause sequence enriched at translation start sites drives transcription dynamics in vivo.

A pause sequence enriched at translation start sites drives transcription dynamics in vivo.
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DOI:
10.1126/science.1251871
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发表时间:
2014-05-30
期刊:
Science (New York, N.Y.)
影响因子:
--
通讯作者:
Weissman JS
Weissman JS
中科院分区:
其他
文献类型:
--
作者:
Larson MH;Mooney RA;Peters JM;Windgassen T;Nayak D;Gross CA;Block SM;Greenleaf WJ;Landick R;Weissman JS

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RNA聚合酶(RNAP)的转录被发挥不同调节作用的停顿打断。虽然在体外已经对个体停顿进行了研究,但体内停顿的决定因素及其在细菌基因组中的分布仍不清楚。使用新生转录测序,我们在大肠杆菌中确定了一个16个核苷酸的共识暂停序列,它解释了已知的调控暂停位点以及体内约20,000个新的暂停位点。在体外的单分子和系综分析表明,这些停顿是由于RNAP/核酸相互作用抑制了下一个核苷酸的加成。来自不同谱系的RNAP也会导致一致的序列暂停,并在大肠杆菌和枯草杆菌中的翻译起始点得到丰富。因此,我们的结果揭示了一种将已知和新发现的暂停事件统一起来的保守机制。
Transcription by RNA polymerase (RNAP) is interrupted by pauses that play diverse regulatory roles. Although individual pauses have been studied in vitro, the determinants of pauses in vivo and their distribution throughout the bacterial genome remain unknown. Using nascent transcript sequencing we identify a 16-nt consensus pause sequence in E. coli that accounts for known regulatory pause sites as well as ~20,000 new in vivo pause sites. In vitro single-molecule and ensemble analyses demonstrate that these pauses result from RNAP/nucleic-acid interactions that inhibit next-nucleotide addition. The consensus sequence also leads to pausing by RNAPs from diverse lineages and is enriched at translation start sites in both E. coli and B. subtilis. Our results thus reveal a conserved mechanism unifying known and newly identified pause events.
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