Double trouble? Gag in conjunction with double insert in HIV protease contributes to reduced darunavir susceptibility Alison Williams 1 , Adriaan Basson 2,3 , Ikechukwu Achilonu 1 , Heini W. Dirr 1 , Lynn Morris 2 and Yasien Sayed 1* 1 Protein Structure-Function Research Unit, School of Molecular and Cell Biology, University of Witwatersrand, Johannesburg, 2050, South Africa 2 Centre for HIV and STIs, National Institute for Communicable Diseases (NICD) of the National Health Laboratory Services (NHLS), Johannesburg, South Africa. 3 HIV Pathogenesis Research Unit, Department of Molecular Medicine and Haematology, Faculty of Health Sciences, University of the Witwatersrand, 7 York Road, Parktown, 2193, Johannesburg, South Africa * Corresponding author: email: yasien.sayed@wits.ac.za, Tel number: +27 11 717 6350 Running title Susceptibility of L38↑N↑L protease to protease inhibitors Abbreviations L38↑N↑L: HIV-1 subtype C protease containing leucine at position 38 followed by a double insertion of asparagine and leucine L38NL: pseudovirion containing L38↑N↑L protease and its related Gag WTGAG L38NL: pseudovirion containing L38↑N↑L protease and a wild-type subtype C Gag Enzymes HIV-1 protease EC 3.4.23.16 Key words HIV protease, Hinge region insertion, Gag, Protease Inhibitor, Darunavir ACCEPTED MANUSCRIPT 10.1042/BCJ20180692 . Please cite using the DOI 10.1042/BCJ20180692 http://dx.doi.org/ up-to-date version is available at encouraged to use the Version of Record that, when published, will replace this version. The most this is an Accepted Manuscript, not the final Version of Record. You are : Biochemical Journal ). http://www.portlandpresspublishing.com/content/open-access-policy#Archiving Archiving Policy of Portland Press ( which the article is published. Archiving of non-open access articles is permitted in accordance with the Use of open access articles is permitted based on the terms of the specific Creative Commons Licence under