A Critical Role of the C-terminal Segment for Allosteric Inhibitor-induced Aberrant Multimerization of HIV-1 Integrase |
| |
Authors: | Nikoloz Shkriabai Venkatasubramanian Dharmarajan Alison Slaughter Jacques J Kessl Ross C Larue Lei Feng James R Fuchs Patrick R Griffin Mamuka Kvaratskhelia |
| |
Institution: | From the ‡Center for Retrovirus Research and College of Pharmacy, The Ohio State University, Columbus, Ohio 43210.;the §Department of Molecular Therapeutics, The Scripps Research Institute, Jupiter, Florida 33458, and ;the ¶Division of Medicinal Chemistry and Pharmacognosy, College of Pharmacy, The Ohio State University, Columbus, Ohio 43210 |
| |
Abstract: | Allosteric HIV-1 integrase (IN) inhibitors (ALLINIs) are a promising class of antiretroviral agents for clinical development. Although ALLINIs promote aberrant IN multimerization and inhibit IN interaction with its cellular cofactor LEDGF/p75 with comparable potencies in vitro, their primary mechanism of action in infected cells is through inducing aberrant multimerization of IN. Crystal structures have shown that ALLINIs bind at the IN catalytic core domain dimer interface and bridge two interacting subunits. However, how these interactions promote higher-order protein multimerization is not clear. Here, we used mass spectrometry-based protein footprinting to monitor surface topology changes in full-length WT and the drug-resistant A128T mutant INs in the presence of ALLINI-2. These experiments have identified protein-protein interactions that extend beyond the direct inhibitor binding site and which lead to aberrant multimerization of WT but not A128T IN. Specifically, we demonstrate that C-terminal residues Lys-264 and Lys-266 play an important role in the inhibitor induced aberrant multimerization of the WT protein. Our findings provide structural clues for exploiting IN multimerization as a new, attractive therapeutic target and are expected to facilitate development of improved inhibitors. |
| |
Keywords: | Drug Resistance Human Immunodeficiency Virus (HIV) Integrase Lentivirus Mass Spectrometry (MS) Pharmacology HIV Integrase Allosteric Inhibitors Drug Resistance Protein Multimerization |
|
|