HIV-1 Gag Binds the Multi-Aminoacyl-tRNA Synthetase Complex via the EPRS Subunit
Host factor tRNAs facilitate the replication of retroviruses such as human immunodeficiency virus type 1 (HIV-1). HIV-1 uses human tRNA<sup>Lys3</sup> as the primer for reverse transcription, and the assembly of HIV-1 structural protein Gag at the plasma membrane (PM) is regulated by mat...
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MDPI AG
2023-02-01
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| Series: | Viruses |
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| Online Access: | https://www.mdpi.com/1999-4915/15/2/474 |
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| author | Danni Jin Yiping Zhu Heidi L. Schubert Stephen P. Goff Karin Musier-Forsyth |
| author_facet | Danni Jin Yiping Zhu Heidi L. Schubert Stephen P. Goff Karin Musier-Forsyth |
| author_sort | Danni Jin |
| collection | DOAJ |
| description | Host factor tRNAs facilitate the replication of retroviruses such as human immunodeficiency virus type 1 (HIV-1). HIV-1 uses human tRNA<sup>Lys3</sup> as the primer for reverse transcription, and the assembly of HIV-1 structural protein Gag at the plasma membrane (PM) is regulated by matrix (MA) domain–tRNA interactions. A large, dynamic multi-aminoacyl-tRNA synthetase complex (MSC) exists in the cytosol and consists of eight aminoacyl-tRNA synthetases (ARSs) and three other cellular proteins. Proteomic studies to identify HIV–host interactions have identified the MSC as part of the HIV-1 Gag and MA interactomes. Here, we confirmed that the MA domain of HIV-1 Gag forms a stable complex with the MSC, mapped the primary interaction site to the linker domain of bi-functional human glutamyl-prolyl-tRNA synthetase (EPRS), and showed that the MA–EPRS interaction was RNA dependent. MA mutations that significantly reduced the EPRS interaction reduced viral infectivity and mapped to MA residues that also interact with phosphatidylinositol-(4,5)-bisphosphate. Overexpression of EPRS or EPRS fragments did not affect susceptibility to HIV-1 infection, and knockdown of EPRS reduced both a control reporter gene and HIV-1 protein translation. EPRS knockdown resulted in decreased progeny virion production, but the decrease could not be attributed to selective effects on virus gene expression, and the specific infectivity of the virions remained unchanged. While the precise function of the Gag–EPRS interaction remains uncertain, we discuss possible effects of the interaction on either virus or host activities. |
| first_indexed | 2024-03-11T08:01:02Z |
| format | Article |
| id | doaj.art-b5891c381b9f4cb7a210349f98c6dc9b |
| institution | Directory Open Access Journal |
| issn | 1999-4915 |
| language | English |
| last_indexed | 2024-03-11T08:01:02Z |
| publishDate | 2023-02-01 |
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| record_format | Article |
| series | Viruses |
| spelling | doaj.art-b5891c381b9f4cb7a210349f98c6dc9b2023-11-16T23:49:29ZengMDPI AGViruses1999-49152023-02-0115247410.3390/v15020474HIV-1 Gag Binds the Multi-Aminoacyl-tRNA Synthetase Complex via the EPRS SubunitDanni Jin0Yiping Zhu1Heidi L. Schubert2Stephen P. Goff3Karin Musier-Forsyth4Department of Chemistry and Biochemistry, Center for Retrovirus Research, Center for RNA Biology, Ohio State University, Columbus, OH 43210, USADepartments of Biochemistry and Molecular Biophysics, and Microbiology and Immunology, Columbia University Medical Center, New York, NY 10032, USADepartment of Biochemistry, University of Utah, Salt Lake City, UT 841122, USADepartments of Biochemistry and Molecular Biophysics, and Microbiology and Immunology, Columbia University Medical Center, New York, NY 10032, USADepartment of Chemistry and Biochemistry, Center for Retrovirus Research, Center for RNA Biology, Ohio State University, Columbus, OH 43210, USAHost factor tRNAs facilitate the replication of retroviruses such as human immunodeficiency virus type 1 (HIV-1). HIV-1 uses human tRNA<sup>Lys3</sup> as the primer for reverse transcription, and the assembly of HIV-1 structural protein Gag at the plasma membrane (PM) is regulated by matrix (MA) domain–tRNA interactions. A large, dynamic multi-aminoacyl-tRNA synthetase complex (MSC) exists in the cytosol and consists of eight aminoacyl-tRNA synthetases (ARSs) and three other cellular proteins. Proteomic studies to identify HIV–host interactions have identified the MSC as part of the HIV-1 Gag and MA interactomes. Here, we confirmed that the MA domain of HIV-1 Gag forms a stable complex with the MSC, mapped the primary interaction site to the linker domain of bi-functional human glutamyl-prolyl-tRNA synthetase (EPRS), and showed that the MA–EPRS interaction was RNA dependent. MA mutations that significantly reduced the EPRS interaction reduced viral infectivity and mapped to MA residues that also interact with phosphatidylinositol-(4,5)-bisphosphate. Overexpression of EPRS or EPRS fragments did not affect susceptibility to HIV-1 infection, and knockdown of EPRS reduced both a control reporter gene and HIV-1 protein translation. EPRS knockdown resulted in decreased progeny virion production, but the decrease could not be attributed to selective effects on virus gene expression, and the specific infectivity of the virions remained unchanged. While the precise function of the Gag–EPRS interaction remains uncertain, we discuss possible effects of the interaction on either virus or host activities.https://www.mdpi.com/1999-4915/15/2/474human immunodeficiency virus type 1multi-aminoacyl-tRNA synthetase complexglutamyl-prolyl tRNA synthetaseHIV-1 MA–tRNA interactionsHIV-dependency factors |
| spellingShingle | Danni Jin Yiping Zhu Heidi L. Schubert Stephen P. Goff Karin Musier-Forsyth HIV-1 Gag Binds the Multi-Aminoacyl-tRNA Synthetase Complex via the EPRS Subunit Viruses human immunodeficiency virus type 1 multi-aminoacyl-tRNA synthetase complex glutamyl-prolyl tRNA synthetase HIV-1 MA–tRNA interactions HIV-dependency factors |
| title | HIV-1 Gag Binds the Multi-Aminoacyl-tRNA Synthetase Complex via the EPRS Subunit |
| title_full | HIV-1 Gag Binds the Multi-Aminoacyl-tRNA Synthetase Complex via the EPRS Subunit |
| title_fullStr | HIV-1 Gag Binds the Multi-Aminoacyl-tRNA Synthetase Complex via the EPRS Subunit |
| title_full_unstemmed | HIV-1 Gag Binds the Multi-Aminoacyl-tRNA Synthetase Complex via the EPRS Subunit |
| title_short | HIV-1 Gag Binds the Multi-Aminoacyl-tRNA Synthetase Complex via the EPRS Subunit |
| title_sort | hiv 1 gag binds the multi aminoacyl trna synthetase complex via the eprs subunit |
| topic | human immunodeficiency virus type 1 multi-aminoacyl-tRNA synthetase complex glutamyl-prolyl tRNA synthetase HIV-1 MA–tRNA interactions HIV-dependency factors |
| url | https://www.mdpi.com/1999-4915/15/2/474 |
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