Abstract
Background The unspliced HIV-1 RNA is used as mRNA for translation of the Gag and Pol proteins and packaged as genomic RNA in the virion. In addition, HIV-1 produces more than 40 differentially spliced transcripts that encode the other viral proteins. Regulation of splicing is essential for the production of all unspliced and spliced viral RNAs. The 5’ leader region of the HIV-1 RNA contains the major 5’ splice site (5’ss) that is used in the production of all spliced RNAs. This splice donor (SD) region can fold a stem-loop structure and the stability of this hairpin may influence splicing by restricting the accessibility of the 5’ss for the splicing machinery. To test this hypothesis, the thermodynamic stability of the SD hairpin was varied through mutation and the effect on RNA splicing was analyzed.
Highlights
The unspliced HIV-1 RNA is used as mRNA for translation of the Gag and Pol proteins and packaged as genomic RNA in the virion
The 5’ leader region of the HIV-1 RNA contains the major 5’ splice site (5’ss) that is used in the production of all spliced RNAs
The thermodynamic stability of the splice donor (SD) hairpin was varied through mutation and the effect on RNA splicing was analyzed
Summary
The unspliced HIV-1 RNA is used as mRNA for translation of the Gag and Pol proteins and packaged as genomic RNA in the virion. Regulation of splicing is essential for the production of all unspliced and spliced viral RNAs. The 5’ leader region of the HIV-1 RNA contains the major 5’ splice site (5’ss) that is used in the production of all spliced RNAs. The 5’ leader region of the HIV-1 RNA contains the major 5’ splice site (5’ss) that is used in the production of all spliced RNAs This splice donor (SD) region can fold a stem-loop structure and the stability of this hairpin may influence splicing by restricting the accessibility of the 5’ss for the splicing machinery. To test this hypothesis, the thermodynamic stability of the SD hairpin was varied through mutation and the effect on RNA splicing was analyzed
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