Imaging the biogenesis of individual HIV-1 virions in live cells

被引:257
作者
Jouvenet, Nolwenn [1 ,2 ]
Bieniasz, Paul D. [1 ,2 ]
Simon, Sanford M. [3 ]
机构
[1] Rockefeller Univ, Aaron Diamond AIDS Res Ctr, New York, NY 10065 USA
[2] Rockefeller Univ, Lab Retrovirol, New York, NY 10065 USA
[3] Rockefeller Univ, Lab Cellular Biophys, New York, NY 10065 USA
关键词
D O I
10.1038/nature06998
中图分类号
O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];
学科分类号
07 ; 0710 ; 09 ;
摘要
Observations of individual virions in live cells have led to the characterization of their attachment, entry and intracellular transport(1). However, the assembly of individual virions has never been observed in real time. Insights into this process have come primarily from biochemical analyses of populations of virions or from microscopic studies of fixed infected cells. Thus, some assembly properties, such as kinetics and location, are either unknown or controversial(2-5). Here we describe quantitatively the genesis of individual virions in real time, from initiation of assembly to budding and release. We studied fluorescently tagged derivatives of Gag, the major structural component of HIV-1-which is sufficient to drive the assembly of virus- like particles(6) - with the use of fluorescence resonance energy transfer, fluorescence recovery after photobleaching and total- internal- reflection fluorescent microscopy in living cells. Virions appeared individually at the plasma membrane, their assembly rate accelerated as Gag protein accumulated in cells, and typically 5 - 6 min was required to complete the assembly of a single virion. These approaches allow a previously unobserved view of the genesis of individual virions and the determination of parameters of viral assembly that are inaccessible with conventional techniques.
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页码:236 / 240
页数:5
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