A CRISPR screen for factors regulating SAMHD1 degradation identifies IFITMs as potent inhibitors of lentiviral particle delivery
Abstract The InterFeron Induced TransMembrane (IFITM) proteins are interferon stimulated genes that restrict many viruses, including HIV-1. SAMHD1 is another restriction factor blocking replication of HIV-1 and other viruses. Some lentiviruses evolved Vpx/Vpr proteins to degrade SAMHD1. However, thi...
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| Format: | Article |
| Language: | English |
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BMC
2018-03-01
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| Series: | Retrovirology |
| Subjects: | |
| Online Access: | http://link.springer.com/article/10.1186/s12977-018-0409-2 |
| _version_ | 1818912401554145280 |
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| author | Ferdinand Roesch Molly OhAinle Michael Emerman |
| author_facet | Ferdinand Roesch Molly OhAinle Michael Emerman |
| author_sort | Ferdinand Roesch |
| collection | DOAJ |
| description | Abstract The InterFeron Induced TransMembrane (IFITM) proteins are interferon stimulated genes that restrict many viruses, including HIV-1. SAMHD1 is another restriction factor blocking replication of HIV-1 and other viruses. Some lentiviruses evolved Vpx/Vpr proteins to degrade SAMHD1. However, this viral antagonism can be perturbed by host mechanisms: a recent study showed that in interferon (IFN) treated THP1 cells, Vpx is unable to degrade SAMHD1. In the present work, we designed an Interferon Stimulated Genes (ISGs)-targeted CRISPR knockout screen in order to identify ISGs regulating this phenotype. We found that IFITM proteins contribute to the IFNα-mediated protection of SAMHD1 by blocking VSV-G-mediated entry of the lentiviral particles delivering Vpx. Consistent with this, IFNα treatment and IFITM expression had no effect when the A-MLV envelope was used for pseudotyping. Using an assay measuring viral entry, we show that IFNα and IFITMs directly block the delivery of Vpx into cells by inhibiting VSV-G viral fusion. Strikingly, the VSV-G envelope was significantly more sensitive to this IFNα entry block and to IFITMs than HIV-1’s natural envelope. This highlights important differences between VSV-G pseudotyped and wild-type HIV-1, in particular relative to the pathways they use for viral entry, suggesting that HIV-1 may have evolved to escape restriction factors blocking entry. |
| first_indexed | 2024-12-19T23:14:01Z |
| format | Article |
| id | doaj.art-b30bf54ef65b4eccb5df11bd3cee48d8 |
| institution | Directory Open Access Journal |
| issn | 1742-4690 |
| language | English |
| last_indexed | 2024-12-19T23:14:01Z |
| publishDate | 2018-03-01 |
| publisher | BMC |
| record_format | Article |
| series | Retrovirology |
| spelling | doaj.art-b30bf54ef65b4eccb5df11bd3cee48d82022-12-21T20:02:09ZengBMCRetrovirology1742-46902018-03-0115111610.1186/s12977-018-0409-2A CRISPR screen for factors regulating SAMHD1 degradation identifies IFITMs as potent inhibitors of lentiviral particle deliveryFerdinand Roesch0Molly OhAinle1Michael Emerman2Divisions of Human Biology and Basic Sciences, Fred Hutchinson Cancer Research CenterDivisions of Human Biology and Basic Sciences, Fred Hutchinson Cancer Research CenterDivisions of Human Biology and Basic Sciences, Fred Hutchinson Cancer Research CenterAbstract The InterFeron Induced TransMembrane (IFITM) proteins are interferon stimulated genes that restrict many viruses, including HIV-1. SAMHD1 is another restriction factor blocking replication of HIV-1 and other viruses. Some lentiviruses evolved Vpx/Vpr proteins to degrade SAMHD1. However, this viral antagonism can be perturbed by host mechanisms: a recent study showed that in interferon (IFN) treated THP1 cells, Vpx is unable to degrade SAMHD1. In the present work, we designed an Interferon Stimulated Genes (ISGs)-targeted CRISPR knockout screen in order to identify ISGs regulating this phenotype. We found that IFITM proteins contribute to the IFNα-mediated protection of SAMHD1 by blocking VSV-G-mediated entry of the lentiviral particles delivering Vpx. Consistent with this, IFNα treatment and IFITM expression had no effect when the A-MLV envelope was used for pseudotyping. Using an assay measuring viral entry, we show that IFNα and IFITMs directly block the delivery of Vpx into cells by inhibiting VSV-G viral fusion. Strikingly, the VSV-G envelope was significantly more sensitive to this IFNα entry block and to IFITMs than HIV-1’s natural envelope. This highlights important differences between VSV-G pseudotyped and wild-type HIV-1, in particular relative to the pathways they use for viral entry, suggesting that HIV-1 may have evolved to escape restriction factors blocking entry.http://link.springer.com/article/10.1186/s12977-018-0409-2HIV-1SAMHD1VpxPseudotypesInterferonIFITM |
| spellingShingle | Ferdinand Roesch Molly OhAinle Michael Emerman A CRISPR screen for factors regulating SAMHD1 degradation identifies IFITMs as potent inhibitors of lentiviral particle delivery Retrovirology HIV-1 SAMHD1 Vpx Pseudotypes Interferon IFITM |
| title | A CRISPR screen for factors regulating SAMHD1 degradation identifies IFITMs as potent inhibitors of lentiviral particle delivery |
| title_full | A CRISPR screen for factors regulating SAMHD1 degradation identifies IFITMs as potent inhibitors of lentiviral particle delivery |
| title_fullStr | A CRISPR screen for factors regulating SAMHD1 degradation identifies IFITMs as potent inhibitors of lentiviral particle delivery |
| title_full_unstemmed | A CRISPR screen for factors regulating SAMHD1 degradation identifies IFITMs as potent inhibitors of lentiviral particle delivery |
| title_short | A CRISPR screen for factors regulating SAMHD1 degradation identifies IFITMs as potent inhibitors of lentiviral particle delivery |
| title_sort | crispr screen for factors regulating samhd1 degradation identifies ifitms as potent inhibitors of lentiviral particle delivery |
| topic | HIV-1 SAMHD1 Vpx Pseudotypes Interferon IFITM |
| url | http://link.springer.com/article/10.1186/s12977-018-0409-2 |
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