Single-molecule measurements of viral ssRNA packaging.

Single-molecule measurements of viral ssRNA packaging.
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DOI:
10.1261/rna.057471.116
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发表时间:
2017-01
期刊:
RNA (New York, N.Y.)
影响因子:
--
通讯作者:
Poranen MM
Poranen MM
中科院分区:
其他
文献类型:
--
作者:
Hanhijärvi KJ;Ziedaite G;Bamford DH;Hæggström E;Poranen MM

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双链RNA(DsRNA)噬菌体的基因组包装已被广泛应用于生化和分子生物学方法的研究。我们采用了现有的其中一种噬菌体的体外包装系统来进行单分子实验。据我们所知,这是首次尝试使用光学镊子研究病毒RNA包装的细节。假单胞菌噬菌体φ6是一种具有三部分基因组的双链RNA病毒。正链(+)单链RNA(SsRNA)基因组前体被包装成预制的原衣壳(PC),在那里合成负链。我们介绍了用φ6 PC对病毒单链RNA包装的单分子测量。我们的数据表明,包装在缓慢和快速阶段断断续续地进行,这可能反映了被包装的单链RNA二级结构的展开差异。虽然平均包装速度较低(0.07-0.54 nm/秒),但在快速包装阶段包装可以达到4.62 nm/秒。
Genome packaging of double-stranded RNA (dsRNA) phages has been widely studied using biochemical and molecular biology methods. We adapted the existing in vitro packaging system of one such phage for single-molecule experimentation. To our knowledge, this is the first attempt to study the details of viral RNA packaging using optical tweezers. Pseudomonas phage φ6 is a dsRNA virus with a tripartite genome. Positive-sense (+) single-stranded RNA (ssRNA) genome precursors are packaged into a preformed procapsid (PC), where negative strands are synthesized. We present single-molecule measurements of the viral ssRNA packaging by the φ6 PC. Our data show that packaging proceeds intermittently in slow and fast phases, which likely reflects differences in the unfolding of the RNA secondary structures of the ssRNA being packaged. Although the mean packaging velocity was relatively low (0.07–0.54 nm/sec), packaging could reach 4.62 nm/sec during the fast packaging phase.
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