TY - JOUR
T1 - Structure-function analyses reveal key molecular determinants of HIV-1 CRF01_AE resistance to the entry inhibitor temsavir
AU - Prévost, Jérémie
AU - Chen, Yaozong
AU - Zhou, Fei
AU - Tolbert, William D.
AU - Gasser, Romain
AU - Medjahed, Halima
AU - Nayrac, Manon
AU - Nguyen, Dung N.
AU - Gottumukkala, Suneetha
AU - Hessell, Ann J.
AU - Rao, Venigalla B.
AU - Pozharski, Edwin
AU - Huang, Rick K.
AU - Matthies, Doreen
AU - Finzi, Andrés
AU - Pazgier, Marzena
N1 - Funding Information:
The authors thank the CRCHUM BSL3 and Flow Cytometry Platforms for technical assistance. We thank Agnes L. Chenine (U.S. Military HIV Research Program) for providing the 92TH023 and CM244 Env expressors. We would like to thank Drs. Di Wu and Grzegorz Piszczek from the Biophysics Core Facility, National Heart, Lung, and Blood Institute NIH, Bethesda, MD, USA for help in initial characterization of temsavir complexes using single-molecule mass photometry. We greatly thank Allison R. Zeher, Abraham J. Morton, Zabrina C. Lang from National Cancer Institute and NIH IRP CryoEM Consortium (NICE) for electron microscopy data collection support. Funding for this study was provided by the National Institute of Health grants (R01 AI148379 and R01 AI150322 to A.F.; R01 AI129769 and R01AI174908 to M.P. and A.F.; and R01 AI129801 to A. H.). Support for this work was also provided by P01 GM56550/AI150471 to A.F. and P01 AI162242 to M.P. and Georgia Tomaras. This work was also supported by CIHR foundation grant 352417 to A.F. and Canada Foundation for Innovation (CFI) grant 41027 (to A.F.), and to F.Z. and D.M. by the Division of Intramural Research of the Eunice Kennedy Shriver National Institute of Child Health and Human Development, NIH (grant NICHD intramural projects Z1A HD008998). This work was partially supported by the ViiV Healthcare grant 219712 to M.P. A.F. is the recipient of Canada Research Chair on Retroviral Entry RCHS0235 950–232424. J.P. is the recipient of a CIHR doctoral fellowship. R.G. was supported by a MITACS Accélération postdoctoral fellowship. CryoEM facilties at IBBR are supported by the University of Maryland Strategic partnership (MPower). Funding bodies had no role in the design, collection, analysis, or interpretation of the data. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript and the contents of this publication are solely the responsibility of the authors.
Funding Information:
The authors thank the CRCHUM BSL3 and Flow Cytometry Platforms for technical assistance. We thank Agnes L. Chenine (U.S. Military HIV Research Program) for providing the 92TH023 and CM244 Env expressors. We would like to thank Drs. Di Wu and Grzegorz Piszczek from the Biophysics Core Facility, National Heart, Lung, and Blood Institute NIH, Bethesda, MD, USA for help in initial characterization of temsavir complexes using single-molecule mass photometry. We greatly thank Allison R. Zeher, Abraham J. Morton, Zabrina C. Lang from National Cancer Institute and NIH IRP CryoEM Consortium (NICE) for electron microscopy data collection support. Funding for this study was provided by the National Institute of Health grants (R01 AI148379 and R01 AI150322 to A.F.; R01 AI129769 and R01AI174908 to M.P. and A.F.; and R01 AI129801 to A. H.). Support for this work was also provided by P01 GM56550/AI150471 to A.F. and P01 AI162242 to M.P. and Georgia Tomaras. This work was also supported by CIHR foundation grant 352417 to A.F. and Canada Foundation for Innovation (CFI) grant 41027 (to A.F.), and to F.Z. and D.M. by the Division of Intramural Research of the Eunice Kennedy Shriver National Institute of Child Health and Human Development, NIH (grant NICHD intramural projects Z1A HD008998). This work was partially supported by the ViiV Healthcare grant 219712 to M.P. A.F. is the recipient of Canada Research Chair on Retroviral Entry RCHS0235 950–232424. J.P. is the recipient of a CIHR doctoral fellowship. R.G. was supported by a MITACS Accélération postdoctoral fellowship. CryoEM facilties at IBBR are supported by the University of Maryland Strategic partnership (MPower). Funding bodies had no role in the design, collection, analysis, or interpretation of the data. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript and the contents of this publication are solely the responsibility of the authors.
Publisher Copyright:
© 2023, Springer Nature Limited.
PY - 2023/12
Y1 - 2023/12
N2 - The HIV-1 entry inhibitor temsavir prevents the viral receptor CD4 (cluster of differentiation 4) from interacting with the envelope glycoprotein (Env) and blocks its conformational changes. To do this, temsavir relies on the presence of a residue with small side chain at position 375 in Env and is unable to neutralize viral strains like CRF01_AE carrying His375. Here we investigate the mechanism of temsavir resistance and show that residue 375 is not the sole determinant of resistance. At least six additional residues within the gp120 inner domain layers, including five distant from the drug-binding pocket, contribute to resistance. A detailed structure-function analysis using engineered viruses and soluble trimer variants reveals that the molecular basis of resistance is mediated by crosstalk between His375 and the inner domain layers. Furthermore, our data confirm that temsavir can adjust its binding mode to accommodate changes in Env conformation, a property that likely contributes to its broad antiviral activity.
AB - The HIV-1 entry inhibitor temsavir prevents the viral receptor CD4 (cluster of differentiation 4) from interacting with the envelope glycoprotein (Env) and blocks its conformational changes. To do this, temsavir relies on the presence of a residue with small side chain at position 375 in Env and is unable to neutralize viral strains like CRF01_AE carrying His375. Here we investigate the mechanism of temsavir resistance and show that residue 375 is not the sole determinant of resistance. At least six additional residues within the gp120 inner domain layers, including five distant from the drug-binding pocket, contribute to resistance. A detailed structure-function analysis using engineered viruses and soluble trimer variants reveals that the molecular basis of resistance is mediated by crosstalk between His375 and the inner domain layers. Furthermore, our data confirm that temsavir can adjust its binding mode to accommodate changes in Env conformation, a property that likely contributes to its broad antiviral activity.
UR - http://www.scopus.com/inward/record.url?scp=85174730149&partnerID=8YFLogxK
U2 - 10.1038/s41467-023-42500-2
DO - 10.1038/s41467-023-42500-2
M3 - Article
AN - SCOPUS:85174730149
SN - 2041-1723
VL - 14
JO - Nature Communications
JF - Nature Communications
IS - 1
M1 - 6710
ER -