Dynamics of HIV-1 Gag Processing as Revealed by Fluorescence Lifetime Imaging Microscopy and Single Virus Tracking
The viral polyprotein Gag plays a central role for HIV-1 assembly, release and maturation. Proteolytic processing of Gag by the viral protease is essential for the structural rearrangements that mark the transition from immature to mature, infectious viruses. The timing and kinetics of Gag processin...
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| Format: | Article |
| Language: | English |
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MDPI AG
2022-02-01
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| Series: | Viruses |
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| Online Access: | https://www.mdpi.com/1999-4915/14/2/340 |
| _version_ | 1797475997696655360 |
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| author | Chen Qian Annica Flemming Barbara Müller Don C. Lamb |
| author_facet | Chen Qian Annica Flemming Barbara Müller Don C. Lamb |
| author_sort | Chen Qian |
| collection | DOAJ |
| description | The viral polyprotein Gag plays a central role for HIV-1 assembly, release and maturation. Proteolytic processing of Gag by the viral protease is essential for the structural rearrangements that mark the transition from immature to mature, infectious viruses. The timing and kinetics of Gag processing are not fully understood. Here, fluorescence lifetime imaging microscopy and single virus tracking are used to follow Gag processing in nascent HIV-1 particles in situ. Using a Gag polyprotein labelled internally with eCFP, we show that proteolytic release of the fluorophore from Gag is accompanied by an increase in its fluorescence lifetime. By tracking nascent virus particles in situ and analyzing the intensity and fluorescence lifetime of individual traces, we detect proteolytic cleavage of eCFP from Gag in a subset (6.5%) of viral particles. This suggests that for the majority of VLPs, Gag processing occurs with a delay after particle assembly. |
| first_indexed | 2024-03-09T20:51:39Z |
| format | Article |
| id | doaj.art-7b22e93cf70c41c8af4d729d0e977920 |
| institution | Directory Open Access Journal |
| issn | 1999-4915 |
| language | English |
| last_indexed | 2024-03-09T20:51:39Z |
| publishDate | 2022-02-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Viruses |
| spelling | doaj.art-7b22e93cf70c41c8af4d729d0e9779202023-11-23T22:31:20ZengMDPI AGViruses1999-49152022-02-0114234010.3390/v14020340Dynamics of HIV-1 Gag Processing as Revealed by Fluorescence Lifetime Imaging Microscopy and Single Virus TrackingChen Qian0Annica Flemming1Barbara Müller2Don C. Lamb3Department of Chemistry, Ludwig Maximilians-Universität München, Butenandtstraße 5-13, 81377 München, GermanyCenter for Integrative Infectious Diseases Research (CIID), Universitätsklinikum Heidelberg, Im Neuenheimer Feld 344, 69120 Heidelberg, GermanyCenter for Integrative Infectious Diseases Research (CIID), Universitätsklinikum Heidelberg, Im Neuenheimer Feld 344, 69120 Heidelberg, GermanyDepartment of Chemistry, Ludwig Maximilians-Universität München, Butenandtstraße 5-13, 81377 München, GermanyThe viral polyprotein Gag plays a central role for HIV-1 assembly, release and maturation. Proteolytic processing of Gag by the viral protease is essential for the structural rearrangements that mark the transition from immature to mature, infectious viruses. The timing and kinetics of Gag processing are not fully understood. Here, fluorescence lifetime imaging microscopy and single virus tracking are used to follow Gag processing in nascent HIV-1 particles in situ. Using a Gag polyprotein labelled internally with eCFP, we show that proteolytic release of the fluorophore from Gag is accompanied by an increase in its fluorescence lifetime. By tracking nascent virus particles in situ and analyzing the intensity and fluorescence lifetime of individual traces, we detect proteolytic cleavage of eCFP from Gag in a subset (6.5%) of viral particles. This suggests that for the majority of VLPs, Gag processing occurs with a delay after particle assembly.https://www.mdpi.com/1999-4915/14/2/340HIVmaturationgag processingfluorescence lifetimesingle virus tracking |
| spellingShingle | Chen Qian Annica Flemming Barbara Müller Don C. Lamb Dynamics of HIV-1 Gag Processing as Revealed by Fluorescence Lifetime Imaging Microscopy and Single Virus Tracking Viruses HIV maturation gag processing fluorescence lifetime single virus tracking |
| title | Dynamics of HIV-1 Gag Processing as Revealed by Fluorescence Lifetime Imaging Microscopy and Single Virus Tracking |
| title_full | Dynamics of HIV-1 Gag Processing as Revealed by Fluorescence Lifetime Imaging Microscopy and Single Virus Tracking |
| title_fullStr | Dynamics of HIV-1 Gag Processing as Revealed by Fluorescence Lifetime Imaging Microscopy and Single Virus Tracking |
| title_full_unstemmed | Dynamics of HIV-1 Gag Processing as Revealed by Fluorescence Lifetime Imaging Microscopy and Single Virus Tracking |
| title_short | Dynamics of HIV-1 Gag Processing as Revealed by Fluorescence Lifetime Imaging Microscopy and Single Virus Tracking |
| title_sort | dynamics of hiv 1 gag processing as revealed by fluorescence lifetime imaging microscopy and single virus tracking |
| topic | HIV maturation gag processing fluorescence lifetime single virus tracking |
| url | https://www.mdpi.com/1999-4915/14/2/340 |
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