Understanding the Assembly process in HIV-1

HIV maturation is a highly regulated process, and viral protease has to cleave precursor of the main structural protein Gag in a sequential manner. Non-sequential proteolytic processing event or excess Gag processing intermediates will render progeny HIV non-infectious. This vulnerability has made ‘virus maturation’ an ideal antiviral target, and it has resulted in the development of two classes of HIV inhibitors aiming at both enzyme (proteases - protease inhibitor) and substrate (Gag - maturation inhibitor, MI). However, a natural polymorphism that resists HIV MIs has limited the clinical application of early generation of these drugs. First generation MIs, such as Bevirimat, delay exposure of the Gag CApsid-Spacer Peptide 1 (CA-SP1) cleavage site to viral protease via stabilizing the SP1 six-helix bundle, which prevents conformational switch of the Gag-derived proteins for cleavage and results in excess p25CA-SP1 processing intermediates in the viral particles. Furthermore, MIs have also been shown to affect the assembly of HIV Gag prior to maturation. This proposal builds upon our pioneer effort in using full-length recombinant Gag to study HIV assembly and maturation. This project will define - The factors that drive Gag to refold as HIV assembly and matures; - How MIs interfere with these processes biochemically and biophysically. Successful completion of this proposal will result in a first comprehensive dataset on the biochemical and biophysical features of HIV assembly and maturation. Information generated from this work will transform our understanding of this process that will improve our design for the next generation of maturation inhibitors for clinical application. This project will require a significant effort in image processing and data analysis, Most of the code is already available in the lab but adjustments and optimization might be required. We use Matlab and Python together with a number of specialistic packages for the analyses. although no prior programming experience is required eagerness to learn is vital.