Link to the University of Pittsburgh Homepage
Link to the University Library System Homepage Link to the Contact Us Form
D-Scholarship @ Pitt Banner and Links to Homepage

Evidence for biphasic uncoating during HIV-1 infection from a novel imaging assay

Xu, H and Franks, T and Gibson, G and Huber, K and Rahm, N and De Castillia, CS and Luban, J and Aiken, C and Watkins, S and Sluis-Cremer, N and Ambrose, Z (2013) Evidence for biphasic uncoating during HIV-1 infection from a novel imaging assay. Retrovirology, 10 (1).

[img]
Preview
 PDF
Published Version
Available under License : See the attached license file.
Download (2MB) | Preview
[img] Plain Text (licence)
Available under License : See the attached license file.
Download (1kB)

Abstract

Background: Uncoating of the HIV-1 core plays a critical role during early post-fusion stages of infection but is poorly understood. Microscopy-based assays are unable to easily distinguish between intact and partially uncoated viral cores.Results: In this study, we used 5-ethynyl uridine (EU) to label viral-associated RNA during HIV production. At early time points after infection with EU-labeled virions, the viral-associated RNA was stained with an EU-specific dye and was detected by confocal microscopy together with viral proteins. We observed that detection of the viral-associated RNA was specific for EU-labeled virions, was detected only after viral fusion with target cells, and occurred after an initial opening of the core. In vitro staining of cores showed that the opening of the core allowed the small molecule dye, but not RNase A or antibodies, inside. Also, staining of the viral-associated RNA, which is co-localized with nucleocapsid, decays over time after viral infection. The decay rate of RNA staining is dependent on capsid (CA) stability, which was altered by CA mutations or a small molecule inducer of HIV-1 uncoating. While the staining of EU-labeled RNA was not affected by inhibition of reverse transcription, the kinetics of core opening of different CA mutants correlated with initiation of reverse transcription. Analysis of the E45A CA mutant suggests that initial core opening is independent of complete capsid disassembly.Conclusions: Taken together, our results establish a novel RNA accessibility-based assay that detects an early event in HIV-1 uncoating and can be used to further define this process. © 2013 Xu et al.; licensee BioMed Central Ltd.

Share

Citation/Export:
Social Networking:
Share |

Details

Item Type: Article
Status: Published
Creators/Authors:
CreatorsEmailPitt UsernameORCID
Xu, H
Franks, T
Gibson, G
Huber, K
Rahm, N
De Castillia, CS
Luban, J
Aiken, C
Watkins, Ssimon.watkins@pitt.eduSWATKINS
Sluis-Cremer, Nnps2@pitt.eduNPS2
Ambrose, Zzaa4@pitt.eduZAA4
Date: 9 July 2013
Date Type: Publication
Journal or Publication Title: Retrovirology
Volume: 10
Number: 1
DOI or Unique Handle: 10.1186/1742-4690-10-70
Schools and Programs: School of Medicine > Cell Biology and Molecular Physiology
School of Medicine > Infectious Diseases and Microbiology
School of Medicine > Medicine
Refereed: Yes
Date Deposited: 07 Oct 2016 15:57
Last Modified: 29 Jan 2019 15:55
URI: http://d-scholarship.pitt.edu/id/eprint/29704

Metrics

Monthly Views for the past 3 years

Plum Analytics

Altmetric.com

Actions (login required)

View Item View Item