HIV-1 Vpr N-terminal tagging affects alternative splicing of the viral genomeReport as inadecuate




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(2016)SCIENTIFIC REPORTS.6. Mark abstract To facilitate studies on Vpr function in replicating HIV-1, we aimed to tag the protein in an infectious virus. First we showed that N-, but not C-terminal HA/FLAG tagging of Vpr protein preserves Vpr cytopathicity. Cloning the tags into proviral DNA however ablated viral production and replication. By construction of additional viral variants we could show this defect was not protein-but RNA-dependent and sequence specific, and characterized by oversplicing of the genomic RNA. Simulation of genomic RNA folding suggested that introduction of the tag sequence induced an alternative folding structure in a region enriched in splice sites and splicing regulatory sequences. In silico predictions identified the HA/His(6)-Vpr tagging in HIV-1 to affect mRNA folding less than HA/FLAG-Vpr tagging. In vitro infectivity and mRNA splice pattern improved but did not reach wild-type values. Thus, sequence-specific insertions may interfere with mRNA splicing, possibly due to altered RNA folding. Our results point to the complexity of viral RNA genome sequence interactions. This should be taken into consideration when designing viral manipulation strategies, for both research as for biological interventions.

Please use this url to cite or link to this publication: http://hdl.handle.net/1854/LU-8159575



Author: Ann Baeyens, Evelien Naessens , Anouk Van Nuffel, Karin Weening , Anne-Marie Reilly, Eva Claeys, Wim Trypsteen , Linos Vandekerckh

Source: https://biblio.ugent.be/publication/8159575



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