Excessive RNA Splicing and Inhibition of HIV-1 Replication Induced by Modified U1 Small Nuclear RNAs
- 15 December 2010
- journal article
- Published by American Society for Microbiology in Journal of Virology
- Vol. 84 (24), 12790-12800
- https://doi.org/10.1128/jvi.01257-10
Abstract
HIV-1 RNA undergoes a complex splicing process whereby over 40 different mRNA species are produced by alternative splicing. In addition, approximately half of the RNA transcripts remain unspliced and either are used to encode Gag and Gag-Pol proteins or are packaged into virions as genomic RNA. It has previously been shown that HIV-1 splicing is regulated by cis elements that bind to cellular factors. These factors either enhance or repress definition of exons that are flanked by the HIV-1 3′ splice sites. Here we report that expression of modified U1 snRNPs with increased affinity to HIV-1 downstream 5′ splice sites and to sequences within the first tat coding exon act to selectively increase splicing at the upstream 3′ splice sites in cotransfected 293T cells. This results in a decrease of unspliced viral RNA levels and an approximately 10-fold decrease in virus production. In addition, excessive splicing of viral RNA is concomitant with a striking reduction in the relative amounts of Gag processing intermediates and products. We also show that T cell lines expressing modified U1 snRNAs exhibit reduced HIV-1 replication. Our results suggest that induction of excessive HIV-1 RNA splicing may be a novel strategy to inhibit virus replication in human patients.Keywords
This publication has 39 references indexed in Scilit:
- Efficient transcription through an intron requires the binding of an Sm-type U1 snRNP with intact stem loop II to the splice donorNucleic Acids Research, 2010
- Regulation of vif mRNA Splicing by Human Immunodeficiency Virus Type 1 Requires 5′ Splice Site D2 and an Exonic Splicing Enhancer To Counteract Cellular Restriction Factor APOBEC3GJournal of Virology, 2009
- Development and characterization of a triple combination gene therapy vector inhibiting HIV‐1 multiplicationThe Journal of Gene Medicine, 2008
- Negative and Positive mRNA Splicing Elements Act Competitively To Regulate Human Immunodeficiency Virus Type 1 Vif Gene ExpressionJournal of Virology, 2008
- Gag-Processing Defect of Human Immunodeficiency Virus Type 1 Integrase E246 and G247 Mutants Is Caused by Activation of an Overlapping 5′ Splice SiteJournal of Virology, 2008
- Diagnostics of pathogenic splicing mutations: does bioinformatics cover all bases?Frontiers in Bioscience-Landmark, 2008
- Use of modified U1 snRNAs to inhibit HIV-1 replicationNucleic Acids Research, 2006
- PANTHER version 6: protein sequence and function evolution data with expanded representation of biological pathwaysNucleic Acids Research, 2006
- U1 snRNP targets an essential splicing factor, U2AF65, to the 3' splice site by a network of interactions spanning the exon.Genes & Development, 1992
- A compensatory base change in U1 snRNA suppresses a 5′ splice site mutationCell, 1986