HUMAN-IMMUNODEFICIENCY-VIRUS TYPE-2 (HIV-2) - PACKAGING SIGNAL AND ASSOCIATED NEGATIVE REGULATORY ELEMENT

Human immunodeficiency virus type 2 (HIV-2)-based retroviral vectors will have several desirable features as vehicles for gene therapy. These include target cell specificity, regulated expression, and attenuated cytopathicity. Such vectors require efficient packaging of RNA into retroviral particles which depends on a cis-acting sequence element called packaging signal or psi site. For most retroviruses, the principal part of this element is located between the major splice donor site and the gag initiator codon (AUG) in the leader sequence. The deletion of the corresponding region of HIV-2 did indeed cause a packaging defect; however, it did not abolish RNA encapsidation and viral infectivity. Additionally, deletions in this region resulted in an increase in intracellular viral RNA and extracellular p27 core antigen. However, only a fraction of the intracellular viral RNA was packaged into mature particles. These effects appeared to be sequence specific as deletion of the sequence elements upstream of the splice donor site did not result in increased viral RNA and proteins. A computer-assisted analysis of the leader sequence of viral RNA shows it to be rich in secondary structure, which was markedly altered in the deletion mutants. Thus, the leader sequence of HIV-2 between the splice donor site and the gag ATG has at least two regulatory functions: one positive, affecting encapsidation, and the other negative, regulating virus expression. Because there is only a limited sequence or structural homology between the corresponding region of HIV-1 and HIV-2, they are likely to differ in their pathways regulating packaging and gene expression.