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Didehydro-Cortistatin A Inhibits HIV-1 by Specifically Binding to the Unstructured Basic Region of Tat

The intrinsically disordered HIV-1 Tat protein binds the viral RNA transactivation response structure (TAR), which recruits transcriptional cofactors, amplifying viral mRNA expression. Limited Tat transactivation correlates with HIV-1 latency. Unfortunately, Tat inhibitors are not clinically availab...

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Autores principales: Mediouni, Sonia, Chinthalapudi, Krishna, Ekka, Mary K., Usui, Ippei, Jablonski, Joseph A., Clementz, Mark A., Mousseau, Guillaume, Nowak, Jason, Macherla, Venkat R., Beverage, Jacob N., Esquenazi, Eduardo, Baran, Phil, de Vera, Ian Mitchelle S., Kojetin, Douglas, Loret, Erwann P., Nettles, Kendall, Maiti, Souvik, Izard, Tina, Valente, Susana T.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: American Society for Microbiology 2019
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6368365/
https://www.ncbi.nlm.nih.gov/pubmed/30723126
http://dx.doi.org/10.1128/mBio.02662-18
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author Mediouni, Sonia
Chinthalapudi, Krishna
Ekka, Mary K.
Usui, Ippei
Jablonski, Joseph A.
Clementz, Mark A.
Mousseau, Guillaume
Nowak, Jason
Macherla, Venkat R.
Beverage, Jacob N.
Esquenazi, Eduardo
Baran, Phil
de Vera, Ian Mitchelle S.
Kojetin, Douglas
Loret, Erwann P.
Nettles, Kendall
Maiti, Souvik
Izard, Tina
Valente, Susana T.
author_facet Mediouni, Sonia
Chinthalapudi, Krishna
Ekka, Mary K.
Usui, Ippei
Jablonski, Joseph A.
Clementz, Mark A.
Mousseau, Guillaume
Nowak, Jason
Macherla, Venkat R.
Beverage, Jacob N.
Esquenazi, Eduardo
Baran, Phil
de Vera, Ian Mitchelle S.
Kojetin, Douglas
Loret, Erwann P.
Nettles, Kendall
Maiti, Souvik
Izard, Tina
Valente, Susana T.
author_sort Mediouni, Sonia
collection PubMed
description The intrinsically disordered HIV-1 Tat protein binds the viral RNA transactivation response structure (TAR), which recruits transcriptional cofactors, amplifying viral mRNA expression. Limited Tat transactivation correlates with HIV-1 latency. Unfortunately, Tat inhibitors are not clinically available. The small molecule didehydro-cortistatin A (dCA) inhibits Tat, locking HIV-1 in persistent latency, blocking viral rebound. We generated chemical derivatives of dCA that rationalized molecular docking of dCA to an active and specific Tat conformer. These revealed the importance of the cycloheptene ring and the isoquinoline nitrogen’s positioning in the interaction with specific residues of Tat’s basic domain. These features are distinct from the ones required for inhibition of cyclin-dependent kinase 8 (CDK8), the only other known ligand of dCA. Besides, we demonstrated that dCA activity on HIV-1 transcription is independent of CDK8. The binding of dCA to Tat with nanomolar affinity alters the local protein environment, rendering Tat more resistant to proteolytic digestion. dCA thus locks a transient conformer of Tat, specifically blocking functions dependent of its basic domain, namely the Tat-TAR interaction; while proteins with similar basic patches are unaffected by dCA. Our results improve our knowledge of the mode of action of dCA and support structure-based design strategies targeting Tat, to help advance development of dCA, as well as novel Tat inhibitors.
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spelling pubmed-63683652019-02-11 Didehydro-Cortistatin A Inhibits HIV-1 by Specifically Binding to the Unstructured Basic Region of Tat Mediouni, Sonia Chinthalapudi, Krishna Ekka, Mary K. Usui, Ippei Jablonski, Joseph A. Clementz, Mark A. Mousseau, Guillaume Nowak, Jason Macherla, Venkat R. Beverage, Jacob N. Esquenazi, Eduardo Baran, Phil de Vera, Ian Mitchelle S. Kojetin, Douglas Loret, Erwann P. Nettles, Kendall Maiti, Souvik Izard, Tina Valente, Susana T. mBio Research Article The intrinsically disordered HIV-1 Tat protein binds the viral RNA transactivation response structure (TAR), which recruits transcriptional cofactors, amplifying viral mRNA expression. Limited Tat transactivation correlates with HIV-1 latency. Unfortunately, Tat inhibitors are not clinically available. The small molecule didehydro-cortistatin A (dCA) inhibits Tat, locking HIV-1 in persistent latency, blocking viral rebound. We generated chemical derivatives of dCA that rationalized molecular docking of dCA to an active and specific Tat conformer. These revealed the importance of the cycloheptene ring and the isoquinoline nitrogen’s positioning in the interaction with specific residues of Tat’s basic domain. These features are distinct from the ones required for inhibition of cyclin-dependent kinase 8 (CDK8), the only other known ligand of dCA. Besides, we demonstrated that dCA activity on HIV-1 transcription is independent of CDK8. The binding of dCA to Tat with nanomolar affinity alters the local protein environment, rendering Tat more resistant to proteolytic digestion. dCA thus locks a transient conformer of Tat, specifically blocking functions dependent of its basic domain, namely the Tat-TAR interaction; while proteins with similar basic patches are unaffected by dCA. Our results improve our knowledge of the mode of action of dCA and support structure-based design strategies targeting Tat, to help advance development of dCA, as well as novel Tat inhibitors. American Society for Microbiology 2019-02-05 /pmc/articles/PMC6368365/ /pubmed/30723126 http://dx.doi.org/10.1128/mBio.02662-18 Text en Copyright © 2019 Mediouni et al. https://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution 4.0 International license (https://creativecommons.org/licenses/by/4.0/) .
spellingShingle Research Article
Mediouni, Sonia
Chinthalapudi, Krishna
Ekka, Mary K.
Usui, Ippei
Jablonski, Joseph A.
Clementz, Mark A.
Mousseau, Guillaume
Nowak, Jason
Macherla, Venkat R.
Beverage, Jacob N.
Esquenazi, Eduardo
Baran, Phil
de Vera, Ian Mitchelle S.
Kojetin, Douglas
Loret, Erwann P.
Nettles, Kendall
Maiti, Souvik
Izard, Tina
Valente, Susana T.
Didehydro-Cortistatin A Inhibits HIV-1 by Specifically Binding to the Unstructured Basic Region of Tat
title Didehydro-Cortistatin A Inhibits HIV-1 by Specifically Binding to the Unstructured Basic Region of Tat
title_full Didehydro-Cortistatin A Inhibits HIV-1 by Specifically Binding to the Unstructured Basic Region of Tat
title_fullStr Didehydro-Cortistatin A Inhibits HIV-1 by Specifically Binding to the Unstructured Basic Region of Tat
title_full_unstemmed Didehydro-Cortistatin A Inhibits HIV-1 by Specifically Binding to the Unstructured Basic Region of Tat
title_short Didehydro-Cortistatin A Inhibits HIV-1 by Specifically Binding to the Unstructured Basic Region of Tat
title_sort didehydro-cortistatin a inhibits hiv-1 by specifically binding to the unstructured basic region of tat
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6368365/
https://www.ncbi.nlm.nih.gov/pubmed/30723126
http://dx.doi.org/10.1128/mBio.02662-18
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