Formation of nuclear CPSF6/CPSF5 biomolecular condensates upon HIV-1 entry into the nucleus is important for productive infection
Abstract The early events of HIV-1 infection involve the transport of the viral core into the nucleus. This event triggers the translocation of CPSF6 from paraspeckles into nuclear speckles forming puncta-like structures. Our investigations revealed that neither HIV-1 integration nor reverse transcr...
| Main Authors: | , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Nature Portfolio
2023-07-01
|
| Series: | Scientific Reports |
| Online Access: | https://doi.org/10.1038/s41598-023-37364-x |
| _version_ | 1797784717213302784 |
|---|---|
| author | Charlotte Luchsinger KyeongEun Lee Gonzalo A. Mardones Vineet N. KewalRamani Felipe Diaz-Griffero |
| author_facet | Charlotte Luchsinger KyeongEun Lee Gonzalo A. Mardones Vineet N. KewalRamani Felipe Diaz-Griffero |
| author_sort | Charlotte Luchsinger |
| collection | DOAJ |
| description | Abstract The early events of HIV-1 infection involve the transport of the viral core into the nucleus. This event triggers the translocation of CPSF6 from paraspeckles into nuclear speckles forming puncta-like structures. Our investigations revealed that neither HIV-1 integration nor reverse transcription is required for the formation of puncta-like structures. Moreover, HIV-1 viruses without viral genome are competent for the induction of CPSF6 puncta-like structures. In agreement with the notion that HIV-1 induced CPSF6 puncta-like structures are biomolecular condensates, we showed that osmotic stress and 1,6-hexanediol induced the disassembly of CPSF6 condensates. Interestingly, replacing the osmotic stress by isotonic media re-assemble CPSF6 condensates in the cytoplasm of the cell. To test whether CPSF6 condensates were important for infection we utilized hypertonic stress, which prevents formation of CPSF6 condensates, during infection. Remarkably, preventing the formation of CPSF6 condensates inhibits the infection of wild type HIV-1 but not of HIV-1 viruses bearing the capsid changes N74D and A77V, which do not form CPSF6 condensates during infection1,2. We also investigated whether the functional partners of CPSF6 are recruited to the condensates upon infection. Our experiments revealed that CPSF5, but not CPSF7, co-localized with CPSF6 upon HIV-1 infection. We found condensates containing CPSF6/CPSF5 in human T cells and human primary macrophages upon HIV-1 infection. Additionally, we observed that the integration cofactor LEDGF/p75 changes distribution upon HIV-1 infection and surrounds the CPSF6/CPSF5 condensates. Overall, our work demonstrated that CPSF6 and CPSF5 are forming biomolecular condensates that are important for infection of wild type HIV-1 viruses. |
| first_indexed | 2024-03-13T00:43:52Z |
| format | Article |
| id | doaj.art-b4643a6316854a37b63672e8d9b91525 |
| institution | Directory Open Access Journal |
| issn | 2045-2322 |
| language | English |
| last_indexed | 2024-03-13T00:43:52Z |
| publishDate | 2023-07-01 |
| publisher | Nature Portfolio |
| record_format | Article |
| series | Scientific Reports |
| spelling | doaj.art-b4643a6316854a37b63672e8d9b915252023-07-09T11:11:15ZengNature PortfolioScientific Reports2045-23222023-07-0113111410.1038/s41598-023-37364-xFormation of nuclear CPSF6/CPSF5 biomolecular condensates upon HIV-1 entry into the nucleus is important for productive infectionCharlotte Luchsinger0KyeongEun Lee1Gonzalo A. Mardones2Vineet N. KewalRamani3Felipe Diaz-Griffero4Department of Microbiology and Immunology, Albert Einstein College of MedicineBasic Research Laboratory, Center for Cancer Research, National Cancer Institute at FrederickFacultad de Medicina Y Ciencia, Universidad San SebastianBasic Research Laboratory, Center for Cancer Research, National Cancer Institute at FrederickDepartment of Microbiology and Immunology, Albert Einstein College of MedicineAbstract The early events of HIV-1 infection involve the transport of the viral core into the nucleus. This event triggers the translocation of CPSF6 from paraspeckles into nuclear speckles forming puncta-like structures. Our investigations revealed that neither HIV-1 integration nor reverse transcription is required for the formation of puncta-like structures. Moreover, HIV-1 viruses without viral genome are competent for the induction of CPSF6 puncta-like structures. In agreement with the notion that HIV-1 induced CPSF6 puncta-like structures are biomolecular condensates, we showed that osmotic stress and 1,6-hexanediol induced the disassembly of CPSF6 condensates. Interestingly, replacing the osmotic stress by isotonic media re-assemble CPSF6 condensates in the cytoplasm of the cell. To test whether CPSF6 condensates were important for infection we utilized hypertonic stress, which prevents formation of CPSF6 condensates, during infection. Remarkably, preventing the formation of CPSF6 condensates inhibits the infection of wild type HIV-1 but not of HIV-1 viruses bearing the capsid changes N74D and A77V, which do not form CPSF6 condensates during infection1,2. We also investigated whether the functional partners of CPSF6 are recruited to the condensates upon infection. Our experiments revealed that CPSF5, but not CPSF7, co-localized with CPSF6 upon HIV-1 infection. We found condensates containing CPSF6/CPSF5 in human T cells and human primary macrophages upon HIV-1 infection. Additionally, we observed that the integration cofactor LEDGF/p75 changes distribution upon HIV-1 infection and surrounds the CPSF6/CPSF5 condensates. Overall, our work demonstrated that CPSF6 and CPSF5 are forming biomolecular condensates that are important for infection of wild type HIV-1 viruses.https://doi.org/10.1038/s41598-023-37364-x |
| spellingShingle | Charlotte Luchsinger KyeongEun Lee Gonzalo A. Mardones Vineet N. KewalRamani Felipe Diaz-Griffero Formation of nuclear CPSF6/CPSF5 biomolecular condensates upon HIV-1 entry into the nucleus is important for productive infection Scientific Reports |
| title | Formation of nuclear CPSF6/CPSF5 biomolecular condensates upon HIV-1 entry into the nucleus is important for productive infection |
| title_full | Formation of nuclear CPSF6/CPSF5 biomolecular condensates upon HIV-1 entry into the nucleus is important for productive infection |
| title_fullStr | Formation of nuclear CPSF6/CPSF5 biomolecular condensates upon HIV-1 entry into the nucleus is important for productive infection |
| title_full_unstemmed | Formation of nuclear CPSF6/CPSF5 biomolecular condensates upon HIV-1 entry into the nucleus is important for productive infection |
| title_short | Formation of nuclear CPSF6/CPSF5 biomolecular condensates upon HIV-1 entry into the nucleus is important for productive infection |
| title_sort | formation of nuclear cpsf6 cpsf5 biomolecular condensates upon hiv 1 entry into the nucleus is important for productive infection |
| url | https://doi.org/10.1038/s41598-023-37364-x |
| work_keys_str_mv | AT charlotteluchsinger formationofnuclearcpsf6cpsf5biomolecularcondensatesuponhiv1entryintothenucleusisimportantforproductiveinfection AT kyeongeunlee formationofnuclearcpsf6cpsf5biomolecularcondensatesuponhiv1entryintothenucleusisimportantforproductiveinfection AT gonzaloamardones formationofnuclearcpsf6cpsf5biomolecularcondensatesuponhiv1entryintothenucleusisimportantforproductiveinfection AT vineetnkewalramani formationofnuclearcpsf6cpsf5biomolecularcondensatesuponhiv1entryintothenucleusisimportantforproductiveinfection AT felipediazgriffero formationofnuclearcpsf6cpsf5biomolecularcondensatesuponhiv1entryintothenucleusisimportantforproductiveinfection |