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Nucleic Acids Research 2005 33(10):3245-3252; doi:10.1093/nar/gki629
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Published online 7 June 2005

© The Author 2005. Published by Oxford University Press. All rights reserved
The online version of this article has been published under an open access model. Users are entitled to use, reproduce, disseminate, or display the open access version of this article for non-commercial purposes provided that: the original authorship is properly and fully attributed; the Journal and Oxford University Press are attributed as the original place of publication with the correct citation details given; if an article is subsequently reproduced or disseminated not in its entirety but only in part or as a derivative work this must be clearly indicated. For commercial re-use, please contact journals.permissions{at}oupjournals.org

Article

Visualizing the dynamic behavior of poliovirus plus-strand RNA in living host cells

Zong-Qiang Cui1,3, Zhi-Ping Zhang1, Xian-En Zhang1,2,*, Ji-Kai Wen1, Ya-Feng Zhou1 and Wei-Hong Xie1

1State Key Laboratory of Virology, Wuhan Institute of Virology, Chinese Academy of Sciences Wuhan 430071, China 2State Key Laboratory of Biomacromolecules, Institute of Biophysics, Chinese Academy of Sciences Beijing 100101, China 3Graduate School, Chinese Academy Sciences Beijing 100039, China

*To whom correspondence should be addressed. Tel: + 86 10 58881508; Fax: +86 10 58881559; Email: Zhangxe{at}mail.most.gov.cn

Received February 16, 2005. Revised April 28, 2005. Accepted May 14, 2005.

Dynamic analysis of viral nucleic acids in host cells is important for understanding virus–host interaction. By labeling endogenous RNA with molecular beacon, we have realized the direct visualization of viral nucleic acids in living host cells and have studied the dynamic behavior of poliovirus plus-strand RNA. Poliovirus plus-strand RNA was observed to display different distribution patterns in living Vero cells at different post-infection time points. Real-time imaging suggested that the translocation of poliovirus plus-strand RNA is a characteristic rearrangement process requiring intact microtubule network of host cells. Confocal-FRAP measurements showed that 49.4 ± 3.2% of the poliovirus plus-strand RNA molecules diffused freely (with a D-value of 9.6 ± 1.6 x 10–10 cm2/s) within their distribution region, while the remaining (50.5 ± 2.9%) were almost immobile and moved very slowly only with change of the RNA distribution region. Under the electron microscope, it was found that virus-induced membrane rearrangement is microtubule-associated in poliovirus-infected Vero cells. These results reveal an entrapment and diffusion mechanism for the movement of poliovirus plus-strand RNA in living mammalian cells, and demonstrate that the mechanism is mainly associated with microtubules and virus-induced membrane structures.


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