Kinetics of HIV-1 capsid uncoating revealed by single-molecule analysis
Uncoating of the metastable HIV-1 capsid is a tightly regulated disassembly process required for release of the viral cDNA prior to nuclear import. To understand the intrinsic capsid disassembly pathway and how it can be modulated, we have developed a single-particle fluorescence microscopy method t...
| Main Authors: | , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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eLife Sciences Publications Ltd
2018-06-01
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| Series: | eLife |
| Subjects: | |
| Online Access: | https://elifesciences.org/articles/34772 |
| _version_ | 1811181138955206656 |
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| author | Chantal L Márquez Derrick Lau James Walsh Vaibhav Shah Conall McGuinness Andrew Wong Anupriya Aggarwal Michael W Parker David A Jacques Stuart Turville Till Böcking |
| author_facet | Chantal L Márquez Derrick Lau James Walsh Vaibhav Shah Conall McGuinness Andrew Wong Anupriya Aggarwal Michael W Parker David A Jacques Stuart Turville Till Böcking |
| author_sort | Chantal L Márquez |
| collection | DOAJ |
| description | Uncoating of the metastable HIV-1 capsid is a tightly regulated disassembly process required for release of the viral cDNA prior to nuclear import. To understand the intrinsic capsid disassembly pathway and how it can be modulated, we have developed a single-particle fluorescence microscopy method to follow the real-time uncoating kinetics of authentic HIV capsids in vitro immediately after permeabilizing the viral membrane. Opening of the first defect in the lattice is the rate-limiting step of uncoating, which is followed by rapid, catastrophic collapse. The capsid-binding inhibitor PF74 accelerates capsid opening but stabilizes the remaining lattice. In contrast, binding of a polyanion to a conserved arginine cluster in the lattice strongly delays initiation of uncoating but does not prevent subsequent lattice disassembly. Our observations suggest that different stages of uncoating can be controlled independently with the interplay between different capsid-binding regulators likely to determine the overall uncoating kinetics. |
| first_indexed | 2024-04-11T09:13:38Z |
| format | Article |
| id | doaj.art-2f6c7c02cb06486a811864100b4ba19b |
| institution | Directory Open Access Journal |
| issn | 2050-084X |
| language | English |
| last_indexed | 2024-04-11T09:13:38Z |
| publishDate | 2018-06-01 |
| publisher | eLife Sciences Publications Ltd |
| record_format | Article |
| series | eLife |
| spelling | doaj.art-2f6c7c02cb06486a811864100b4ba19b2022-12-22T04:32:25ZengeLife Sciences Publications LtdeLife2050-084X2018-06-01710.7554/eLife.34772Kinetics of HIV-1 capsid uncoating revealed by single-molecule analysisChantal L Márquez0Derrick Lau1James Walsh2Vaibhav Shah3Conall McGuinness4Andrew Wong5Anupriya Aggarwal6Michael W Parker7https://orcid.org/0000-0002-3101-1138David A Jacques8https://orcid.org/0000-0002-6426-4510Stuart Turville9Till Böcking10https://orcid.org/0000-0003-1165-3122EMBL Australia Node in Single Molecule Science, School of Medical Sciences, UNSW, Sydney, Australia; ARC Centre of Excellence in Advanced Molecular Imaging, UNSW, Sydney, AustraliaEMBL Australia Node in Single Molecule Science, School of Medical Sciences, UNSW, Sydney, Australia; ARC Centre of Excellence in Advanced Molecular Imaging, UNSW, Sydney, AustraliaEMBL Australia Node in Single Molecule Science, School of Medical Sciences, UNSW, Sydney, Australia; ARC Centre of Excellence in Advanced Molecular Imaging, UNSW, Sydney, AustraliaEMBL Australia Node in Single Molecule Science, School of Medical Sciences, UNSW, Sydney, Australia; ARC Centre of Excellence in Advanced Molecular Imaging, UNSW, Sydney, AustraliaEMBL Australia Node in Single Molecule Science, School of Medical Sciences, UNSW, Sydney, Australia; ARC Centre of Excellence in Advanced Molecular Imaging, UNSW, Sydney, AustraliaThe Kirby Institute, Sydney, AustraliaThe Kirby Institute, Sydney, AustraliaAustralian Cancer Research Foundation Rational Drug Discovery Centre, St. Vincent’s Institute of Medical Research, Melbourne, Australia; Department of Biochemistry and Molecular Biology, Bio21 Molecular Science and Biotechnology Institute, University of Melbourne, Melbourne, AustraliaEMBL Australia Node in Single Molecule Science, School of Medical Sciences, UNSW, Sydney, AustraliaThe Kirby Institute, Sydney, AustraliaEMBL Australia Node in Single Molecule Science, School of Medical Sciences, UNSW, Sydney, Australia; ARC Centre of Excellence in Advanced Molecular Imaging, UNSW, Sydney, AustraliaUncoating of the metastable HIV-1 capsid is a tightly regulated disassembly process required for release of the viral cDNA prior to nuclear import. To understand the intrinsic capsid disassembly pathway and how it can be modulated, we have developed a single-particle fluorescence microscopy method to follow the real-time uncoating kinetics of authentic HIV capsids in vitro immediately after permeabilizing the viral membrane. Opening of the first defect in the lattice is the rate-limiting step of uncoating, which is followed by rapid, catastrophic collapse. The capsid-binding inhibitor PF74 accelerates capsid opening but stabilizes the remaining lattice. In contrast, binding of a polyanion to a conserved arginine cluster in the lattice strongly delays initiation of uncoating but does not prevent subsequent lattice disassembly. Our observations suggest that different stages of uncoating can be controlled independently with the interplay between different capsid-binding regulators likely to determine the overall uncoating kinetics.https://elifesciences.org/articles/34772HIV-1capsid disassemblyfluorescence miscoscopysingle-molecule |
| spellingShingle | Chantal L Márquez Derrick Lau James Walsh Vaibhav Shah Conall McGuinness Andrew Wong Anupriya Aggarwal Michael W Parker David A Jacques Stuart Turville Till Böcking Kinetics of HIV-1 capsid uncoating revealed by single-molecule analysis eLife HIV-1 capsid disassembly fluorescence miscoscopy single-molecule |
| title | Kinetics of HIV-1 capsid uncoating revealed by single-molecule analysis |
| title_full | Kinetics of HIV-1 capsid uncoating revealed by single-molecule analysis |
| title_fullStr | Kinetics of HIV-1 capsid uncoating revealed by single-molecule analysis |
| title_full_unstemmed | Kinetics of HIV-1 capsid uncoating revealed by single-molecule analysis |
| title_short | Kinetics of HIV-1 capsid uncoating revealed by single-molecule analysis |
| title_sort | kinetics of hiv 1 capsid uncoating revealed by single molecule analysis |
| topic | HIV-1 capsid disassembly fluorescence miscoscopy single-molecule |
| url | https://elifesciences.org/articles/34772 |
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