DDX17 Specifically, and Independently of DDX5, Controls Use of the HIV A4/5 Splice Acceptor Cluster and Is Essential for Efficient Replication of HIV.
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Authors
Sithole, Nyaradzai
Williams, Claire A
Vaughan, Aisling M
Kenyon, Julia C
Lever, Andrew ML
Publication Date
2018-09-14Journal Title
J Mol Biol
ISSN
0022-2836
Publisher
Elsevier BV
Volume
430
Issue
18 Pt B
Pages
3111-3128
Language
eng
Type
Article
Physical Medium
Print-Electronic
Metadata
Show full item recordCitation
Sithole, N., Williams, C. A., Vaughan, A. M., Kenyon, J. C., & Lever, A. M. (2018). DDX17 Specifically, and Independently of DDX5, Controls Use of the HIV A4/5 Splice Acceptor Cluster and Is Essential for Efficient Replication of HIV.. J Mol Biol, 430 (18 Pt B), 3111-3128. https://doi.org/10.1016/j.jmb.2018.06.052
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
Cells, Cultured, Cell Line, Tumor, Humans, HIV-1, HIV Infections, RNA Splice Sites, Gene Expression Regulation, Viral, Alternative Splicing, Amino Acid Motifs, Protein Binding, DEAD-box RNA Helicases, Protein Interaction Domains and Motifs, Gene Knockdown Techniques
Sponsorship
Wellcome Trust , Cambridge Biomedical Research Centre and Clinical Academic Reserve
Funder references
Wellcome Trust (097223/Z/11/Z)
Identifiers
External DOI: https://doi.org/10.1016/j.jmb.2018.06.052
This record's URL: https://www.repository.cam.ac.uk/handle/1810/280044
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