A novel splice donor site in the gag-pol gene is required for HIV-1 RNA stability

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  • Martin Lützelberger, Denmark
  • Line S Reinert
  • Atze T Das, Department of Human Retrovirology, Academic Medical Center, University of Amsterdam, Netherlands
  • Ben Berkhout, Department of Human Retrovirology, Academic Medical Center, University of Amsterdam, Netherlands
  • Jørgen Kjems
Productive infection and successful replication of human immunodeficiency virus 1 (HIV-1) requires the balanced expression of all viral genes. This is achieved by a combination of alternative splicing events and regulated nuclear export of viral RNA. Because viral splicing is incomplete and intron-containing RNAs must be exported from the nucleus where they are normally retained, it must be ensured that the unspliced HIV-1 RNA is actively exported from the nucleus and protected from degradation by processes such as nonsense-mediated decay. Here we report the identification of a novel 178-nt-long exon located in the gag-pol gene of HIV-1 and its inclusion in at least two different mRNA species. Although efficiently spliced in vitro, this exon appears to be tightly repressed and infrequently used in vivo. The splicing is activated or repressed in vitro by the splicing factors ASF/SF2 and heterogeneous nuclear ribonucleoprotein A1, respectively, suggesting that splicing is controlled by these factors. Interestingly, mutations in the 5'-splice site resulted in a dramatic reduction in the steady-state level of HIV-1 RNA, and this effect was partially reversed by expression of U1 small nuclear RNA harboring the compensatory mutation. This implies that U1 small nuclear RNA binding to optimal but non-functional splice sites might have a role in protecting unspliced HIV-1 mRNA from degradation.
Original languageEnglish
JournalJournal of Biological Chemistry
Volume281
Issue27
Pages (from-to)18644-18651
Number of pages8
ISSN0021-9258
DOIs
Publication statusPublished - 7 Jul 2006

    Research areas

  • Amino Acid Sequence, Base Sequence, Cell Line, Exons, Fusion Proteins, gag-pol, Genome, Viral, HIV-1, Humans, Molecular Sequence Data, Mutation, RNA Splice Sites, RNA Splicing, RNA, Messenger, RNA, Viral, Virus Replication

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