@article{mbs:/content/journal/jgv/10.1099/vir.0.045492-0, author = "Capel, Elena and Martrus, Glòria and Parera, Mariona and Clotet, Bonaventura and Martínez, Miguel Angel", title = "Evolution of the human immunodeficiency virus type 1 protease: effects on viral replication capacity and protease robustness", journal= "Journal of General Virology", year = "2012", volume = "93", number = "12", pages = "2625-2634", doi = "https://doi.org/10.1099/vir.0.045492-0", url = "https://www.microbiologyresearch.org/content/journal/jgv/10.1099/vir.0.045492-0", publisher = "Microbiology Society", issn = "1465-2099", type = "Journal Article", abstract = "The rapid spread of human immunodeficiency virus type 1 (HIV-1) in humans has been accompanied by continuous extensive genetic diversification of the virus. The aim of this study was to investigate the impact of HIV-1 diversification on HIV-1 replication capacity (RC) and mutational robustness. Thirty-three HIV-1 protease sequences were amplified from three groups of viruses: two naïve sample groups isolated 15 years apart plus a third group of protease inhibitor-(PI) resistant samples. The amplified proteases were recombined with an HXB2 infectious clone and RC was determined in MT-4 cells. RC was also measured in these three groups after random mutagenesis in vitro using error-prone PCR. No significant RC differences were observed between recombinant viruses from either early or recent naïve isolates (P = 0.5729), even though the proteases from the recent isolates had significantly lower sequence conservation scores compared with a subtype B ancestral sequence (P<0.0001). Randomly mutated recombinant viruses from the three groups exhibited significantly lower RC values than the corresponding wild-type viruses (P<0.0001). There was no significant difference regarding viral infectivity reduction between viruses carrying randomly mutated naïve proteases from early or recent sample isolates (P = 0.8035). Interestingly, a significantly greater loss of RC was observed in the PI-resistant protease group (P = 0.0400). These results demonstrate that protease sequence diversification has not affected HIV-1 RC or protease robustness and indicate that proteases carrying PI resistance substitutions are less robust than naïve proteases.", }