Abrogating ALIX Interactions Results in Stuttering of the ESCRT Machinery
Endosomal sorting complexes required for transport (ESCRT) proteins assemble on budding cellular membranes and catalyze their fission. Using live imaging of HIV virions budding from cells, we followed recruitment of ESCRT proteins ALIX, CHMP4B and VPS4. We report that the ESCRT proteins transiently...
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| Format: | Article |
| Language: | English |
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MDPI AG
2020-09-01
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| Series: | Viruses |
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| Online Access: | https://www.mdpi.com/1999-4915/12/9/1032 |
| _version_ | 1827706071142105088 |
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| author | Shilpa Gupta Mourad Bendjennat Saveez Saffarian |
| author_facet | Shilpa Gupta Mourad Bendjennat Saveez Saffarian |
| author_sort | Shilpa Gupta |
| collection | DOAJ |
| description | Endosomal sorting complexes required for transport (ESCRT) proteins assemble on budding cellular membranes and catalyze their fission. Using live imaging of HIV virions budding from cells, we followed recruitment of ESCRT proteins ALIX, CHMP4B and VPS4. We report that the ESCRT proteins transiently co-localize with virions after completion of virion assembly for durations of 45 ± 30 s. We show that mutagenizing the YP domain of Gag which is the primary ALIX binding site or depleting ALIX from cells results in multiple recruitments of the full ESCRT machinery on the same virion (referred to as stuttering where the number of recruitments to the same virion >3). The stuttering recruitments are approximately 4 ± 3 min apart and have the same stoichiometry of ESCRTs and same residence time (45 ± 30 s) as the single recruitments in wild type interactions. Our observations suggest a role for ALIX during fission and question the linear model of ESCRT recruitment, suggesting instead a more complex co-assembly model. |
| first_indexed | 2024-03-10T16:17:21Z |
| format | Article |
| id | doaj.art-319f084844124dbb9cba3cb9b1b5ef6d |
| institution | Directory Open Access Journal |
| issn | 1999-4915 |
| language | English |
| last_indexed | 2024-03-10T16:17:21Z |
| publishDate | 2020-09-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Viruses |
| spelling | doaj.art-319f084844124dbb9cba3cb9b1b5ef6d2023-11-20T13:58:07ZengMDPI AGViruses1999-49152020-09-01129103210.3390/v12091032Abrogating ALIX Interactions Results in Stuttering of the ESCRT MachineryShilpa Gupta0Mourad Bendjennat1Saveez Saffarian2Center for Cell and Genome Sciences, University of Utah, Salt Lake City, UT 84112, USACenter for Cell and Genome Sciences, University of Utah, Salt Lake City, UT 84112, USACenter for Cell and Genome Sciences, University of Utah, Salt Lake City, UT 84112, USAEndosomal sorting complexes required for transport (ESCRT) proteins assemble on budding cellular membranes and catalyze their fission. Using live imaging of HIV virions budding from cells, we followed recruitment of ESCRT proteins ALIX, CHMP4B and VPS4. We report that the ESCRT proteins transiently co-localize with virions after completion of virion assembly for durations of 45 ± 30 s. We show that mutagenizing the YP domain of Gag which is the primary ALIX binding site or depleting ALIX from cells results in multiple recruitments of the full ESCRT machinery on the same virion (referred to as stuttering where the number of recruitments to the same virion >3). The stuttering recruitments are approximately 4 ± 3 min apart and have the same stoichiometry of ESCRTs and same residence time (45 ± 30 s) as the single recruitments in wild type interactions. Our observations suggest a role for ALIX during fission and question the linear model of ESCRT recruitment, suggesting instead a more complex co-assembly model.https://www.mdpi.com/1999-4915/12/9/1032ESCRTstutteringHIVALIXCHMP4BVPS4 |
| spellingShingle | Shilpa Gupta Mourad Bendjennat Saveez Saffarian Abrogating ALIX Interactions Results in Stuttering of the ESCRT Machinery Viruses ESCRT stuttering HIV ALIX CHMP4B VPS4 |
| title | Abrogating ALIX Interactions Results in Stuttering of the ESCRT Machinery |
| title_full | Abrogating ALIX Interactions Results in Stuttering of the ESCRT Machinery |
| title_fullStr | Abrogating ALIX Interactions Results in Stuttering of the ESCRT Machinery |
| title_full_unstemmed | Abrogating ALIX Interactions Results in Stuttering of the ESCRT Machinery |
| title_short | Abrogating ALIX Interactions Results in Stuttering of the ESCRT Machinery |
| title_sort | abrogating alix interactions results in stuttering of the escrt machinery |
| topic | ESCRT stuttering HIV ALIX CHMP4B VPS4 |
| url | https://www.mdpi.com/1999-4915/12/9/1032 |
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