The HIV-1 capsid-binding host factor CPSF6 is post-transcriptionally regulated by the cellular microRNA miR-125b
| Autor: | Gregory A. Sowd, Jui Pandhare, Joseph Holland, Chandravanu Dash, Muthukumar Balasubramaniam, Adrian Padron, Alan Engelman, Evan Chaudhuri, Fernando Villalta, Sabyasachi Dash |
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| Rok vydání: | 2020 |
| Předmět: |
0301 basic medicine
Virus Integration In silico HIV Infections RNA-binding protein Cleavage and polyadenylation specificity factor Biology Biochemistry 03 medical and health sciences Capsid microRNA Humans Gene Regulation Luciferase 3' Untranslated Regions Molecular Biology Post-transcriptional regulation mRNA Cleavage and Polyadenylation Factors Gene knockdown Binding Sites 030102 biochemistry & molecular biology Three prime untranslated region Cell Biology Cell biology MicroRNAs 030104 developmental biology Mutation HIV-1 |
| Zdroj: | J Biol Chem |
| ISSN: | 0021-9258 |
| DOI: | 10.1074/jbc.ra119.010534 |
| Popis: | Cleavage and polyadenylation specificity factor 6 (CPSF6) is a cellular protein involved in mRNA processing. Emerging evidence suggests that CPSF6 also plays key roles in HIV-1 infection, specifically during nuclear import and integration targeting. However, the cellular and molecular mechanisms that regulate CPSF6 expression are largely unknown. In this study, we report a post-transcriptional mechanism that regulates CPSF6 via the cellular microRNA miR-125b. An in silico analysis revealed that the 3′UTR of CPSF6 contains a miR-125b–binding site that is conserved across several mammalian species. Because miRNAs repress protein expression, we tested the effects of miR-125b expression on CPSF6 levels in miR-125b knockdown and over-expression experiments, revealing that miR-125b and CPSF6 levels are inversely correlated. To determine whether miR-125b post-transcriptionally regulates CPSF6, we introduced the 3′UTR of CPSF6 mRNA into a luciferase reporter and found that miR-125b negatively regulates CPSF6 3′UTR-driven luciferase activity. Accordingly, mutations in the miR-125b seed sequence abrogated the regulatory effect of the miRNA on the CPSF6 3′UTR. Finally, pulldown experiments demonstrated that miR-125b physically interacts with CPSF6 3′UTR. Interestingly, HIV-1 infection down-regulated miR-125b expression concurrent with up-regulation of CPSF6. Notably, miR-125b down-regulation in infected cells was not due to reduced pri-miRNA or pre-miRNA levels. However, miR-125b down-regulation depended on HIV-1 reverse transcription but not viral DNA integration. These findings establish a post-transcriptional mechanism that controls CPSF6 expression and highlight a novel function of miR-125b during HIV-host interaction. |
| Databáze: | OpenAIRE |
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