DDX17 Specifically, and Independently of DDX5, Controls Use of the HIV A4/5 Splice Acceptor Cluster and Is Essential for Efficient Replication of HIV

https://doi.org/10.1016/j.jmb.2018.06.052Get rights and content
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Highlights

  • Regulation of HIV splicing is significantly dependent on host cellular factors.

  • DDX17 interacts with splicing factors and is essential for efficient HIV-1 splicing.

  • DDX17 utilizes distinct RNA/ATPase motifs and acts independently of DDX5.

  • DDX17 C-terminal domain is critical for the role of DDX17 in HIV-1 replication.

Abstract

HIV splicing involves five splice donor and eight splice acceptor sequences which, together with cryptic splice sites, generate over 100 mRNA species. Ninety percent of both partially spliced and fully spliced transcripts utilize the intrinsically weak A4/A5 3′ splice site cluster. We show that DDX17, but not its close paralog DDX5, specifically controls the usage of this splice acceptor group. In its absence, production of the viral envelope protein and other regulatory and accessory proteins is grossly reduced, while Vif, which uses the A1 splice acceptor, is unaffected. This is associated with a profound decrease in viral export from the cell. Loss of Vpu expression causing upregulation of cellular Tetherin compounds the phenotype. DDX17 utilizes distinct RNA binding motifs for its role in efficient HIV replication, and we identify RNA binding motifs essential for its role, while the Walker A, Walker B (DEAD), Q motif and the glycine doublet motif are all dispensable. We show that DDX17 interacts with SRSF1/SF2 and the heterodimeric auxiliary factor U2AF65/35, which are essential splicing factors in the generation of Rev and Env/Vpu transcripts.

Keywords

DDX17/HIV-1
splicing factors
A4/A5 3′ splice site cluster

Abbreviations

ESEs
exonic splicing enhancers
DAPI
4′,6-diamidino-2-phenlindole

Cited by (0)

Current Address: Specialist Virology Centre, Department of Microbiology, Norfolk and Norwich University Hospitals, Norwich, UK.

Current address: Division of Virology, Department of Medicine, St Mary’s campus, Imperial College London, Norfolk Place, W2 1PG, London, UK.