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. 2007 Oct 24:4:77.
doi: 10.1186/1742-4690-4-77.

Isolated HIV-1 core is active for reverse transcription

David Warrilow  1 , Deborah Stenzel, David Harrich
Affiliations

Isolated HIV-1 core is active for reverse transcription

David Warrilow et al. Retrovirology. .

Abstract

Whether purified HIV-1 virion cores are capable of reverse transcription or require uncoating to be activated is currently controversial. To address this question we purified cores from a virus culture and tested for the ability to generate authentic reverse transcription products. A dense fraction (approximately 1.28 g/ml) prepared without detergent, possibly derived from disrupted virions, was found to naturally occur as a minor sub-fraction in our preparations. Core-like particles were identified in this active fraction by electron microscopy. We are the first to report the detection of authentic strong-stop, first-strand transfer and full-length minus strand products in this core fraction without requirement for an uncoating activity.

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Figures

Figure 1
Figure 1
Analysis of core fractions. (A) Endogenous reverse transcriptase activity: strong-stop (squares), first-strand transfer (diamonds) and full-length targets (triangles) are shown. (B) p24 ELISA on fractions; inset shows the density of fractions calculated from weight (fractions 3–9 only are shown). Viral proteins were detected in HIV-1NL4.3 equilibrium gradient fractions 1–9 by western analysis using (C) anti-HIV-1 polyclonal antibody, (D) colorimetric reverse transcriptase ELISA using homoploymeric template (fractions 5–9 only are shown), and (E) anti-gp41 antibody. (F) Negative staining transmission electron microscopy of dense fractions showing four representative core-like structures. 100,000× magnification; bar indicates 50 nm. Please note, the fractions shown in A and B are from a separate preparation to those in C-E and hence fraction numbers do not directly correspond [see Additional file 1 for complete methods].
See this image and copyright information in PMC

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