Structural model of the complete poly(A) region of HIV-1 pre-mRNA

In the HIV-1 retrovirus, identical sequences encompassing the AAUAAA hexamer and the U/GU-rich downstream sequence element (DSE) that compose the core poly(A) site are present at both the 5 and 3 ends of the HIV-1 pre-mRNA. The AAUAAA hexamer is partly occluded by base pairing in the upper part of a semi-stable polyA hairpin. This sets the stage for regulation of HIV-1 polyadenylation, which involves reaction suppression at the 5 end and its stimulation at the 3 end. Efficient utilization of the 3 core poly(A) site is promoted by major and minor upstream sequence elements (USEs) which are uniquely present at the 3 end of the HIV-1 transcript. The structures of the HIV-1 5 and 3 poly(A) sites are defined by overall architecture of complete 5 and 3 untranslated terminal regions (UTRs). To our knowledge, there is still no structural model of a complete 3 UTR of the HIV-1 pre-mRNA and complete 3 poly(A) region including the USEs except the fact that the polyA and transactivation response (TAR) hairpins are present at both ends of the HIV-1 pre-mRNA. In this work, we predicted a secondary structure of the 3 UTR of HIV-1 pre-mRNA based on our observation that the minor USEs are located in a region with a high potential to form G-quadruplex structures. We first present structural models for the major USE, complete 3 poly(A) region, and almost entire 3 UTR of HIV-1 pre-mRNA. Our models are built based on the mfold and UNAFold secondary structure prediction of these regions for about 1500 HIV-1 isolates of different subtypes and recombinant forms. We have demonstrated that these models are valid for most of the HIV-1 isolates studied. The proposed models include the known TAR and polyA hairpins and new structural elements containing the U-rich tract of the major USE and U/GU-rich DSE which are fully exposed and accessible to the polyadenylation machinery, which confirms the functional competence of our models.