Publication Date:
2008-05-27
Description:
Observations of individual virions in live cells have led to the characterization of their attachment, entry and intracellular transport. 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. 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-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.〈br /〉〈br /〉〈a href="https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2708942/" target="_blank"〉〈img src="https://static.pubmed.gov/portal/portal3rc.fcgi/4089621/img/3977009" border="0"〉〈/a〉 〈a href="https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2708942/" target="_blank"〉This paper as free author manuscript - peer-reviewed and accepted for publication〈/a〉〈br /〉〈br /〉〈span class="detail_caption"〉Notes: 〈/span〉Jouvenet, Nolwenn -- Bieniasz, Paul D -- Simon, Sanford M -- P20 GM072015/GM/NIGMS NIH HHS/ -- P20 GM072015-01/GM/NIGMS NIH HHS/ -- P20 GM072015-02/GM/NIGMS NIH HHS/ -- P20 GM072015-02S1/GM/NIGMS NIH HHS/ -- P20 GM072015-03/GM/NIGMS NIH HHS/ -- P20 GM072015-04/GM/NIGMS NIH HHS/ -- P20 GM072015-04S1/GM/NIGMS NIH HHS/ -- R01 AI089844/AI/NIAID NIH HHS/ -- R01 GM087977/GM/NIGMS NIH HHS/ -- England -- Nature. 2008 Jul 10;454(7201):236-40. doi: 10.1038/nature06998. Epub 2008 May 25.〈br /〉〈span class="detail_caption"〉Author address: 〈/span〉Aaron Diamond AIDS Research Center, The Rockefeller University, New York, New York 10065, USA.〈br /〉〈span class="detail_caption"〉Record origin:〈/span〉 〈a href="http://www.ncbi.nlm.nih.gov/pubmed/18500329" target="_blank"〉PubMed〈/a〉
Keywords:
Cell Membrane/metabolism
;
Cell Survival
;
Fluorescence Recovery After Photobleaching
;
Fluorescence Resonance Energy Transfer
;
Fluorescent Dyes/*analysis
;
HIV-1/genetics/*growth & development/metabolism
;
HeLa Cells
;
Humans
;
Kinetics
;
Microscopy, Fluorescence
;
Time Factors
;
Virion/*growth & development/metabolism
;
*Virus Replication
;
gag Gene Products, Human Immunodeficiency Virus/genetics/metabolism
Print ISSN:
0028-0836
Electronic ISSN:
1476-4687
Topics:
Biology
,
Chemistry and Pharmacology
,
Medicine
,
Natural Sciences in General
,
Physics
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