The Virus Entry Pathway Determines Sensitivity to the Antiviral Peptide TAT-I24

The peptide TAT-I24, a fusion of the TAT peptide (amino acids 48–60) and the 9-mer peptide I24, has been previously shown to neutralize several double-stranded (ds) DNA viruses in vitro. We have now extended the testing to potentially sensitive RNA viruses and analyzed the antiviral effect of the pe...

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Main Authors: Eva Kicker, Antonio Kouros, Kurt Zatloukal, Hanna Harant
Format: Article
Language:English
Published: MDPI AG 2025-03-01
Series:Viruses
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Online Access:https://www.mdpi.com/1999-4915/17/4/458
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author Eva Kicker
Antonio Kouros
Kurt Zatloukal
Hanna Harant
author_facet Eva Kicker
Antonio Kouros
Kurt Zatloukal
Hanna Harant
author_sort Eva Kicker
collection DOAJ
description The peptide TAT-I24, a fusion of the TAT peptide (amino acids 48–60) and the 9-mer peptide I24, has been previously shown to neutralize several double-stranded (ds) DNA viruses in vitro. We have now extended the testing to potentially sensitive RNA viruses and analyzed the antiviral effect of the peptide against Severe Acute Respiratory Syndrome Coronavirus-2 (SARS-CoV-2). In Vero E6 cells, TAT-I24 neutralized the human 2019-nCoV isolate (Wuhan variant) in a dose-dependent manner, while it was unable to neutralize two SARS-CoV-2 variants of concern, Delta and Omicron. Moreover, TAT-I24 could not significantly neutralize any of the SARS-CoV-2 variants in the human lung carcinoma cell line Calu-3, which provides an alternative entry route for SARS-CoV-2 by direct membrane fusion. Therefore, a possible dependence on virus uptake by endocytosis was investigated by exposing Vero E6 cells to chloroquine (CQ), an inhibitor of endosomal acidification. The Wuhan variant was highly sensitive to inhibition by CQ, an effect which was further enhanced by TAT-I24, while the Delta variant was less sensitive to inhibition by higher concentrations of CQ compared to the Wuhan variant. The microscopic analysis of COS-7 cells using a rhodamine-labeled TAT-I24 (Rho-TAT-I24) showed the endosomal localization of fluorescent TAT-I24 and co-localization with transfected GFP-Rab14 but not GFP-Rab5. As these proteins are found in distinct endosomal pathways, our results indicate that the virus entry pathway determines sensitivity to the peptide.
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spelling doaj-art-d0788a2cff9e46ab8f11a3bf5bfa72e72025-08-20T03:13:33ZengMDPI AGViruses1999-49152025-03-0117445810.3390/v17040458The Virus Entry Pathway Determines Sensitivity to the Antiviral Peptide TAT-I24Eva Kicker0Antonio Kouros1Kurt Zatloukal2Hanna Harant3Diagnostic and Research Center for Molecular Biomedicine, Diagnostic and Research Institute of Pathology, Medical University Graz, Neue Stiftingtalstraße 6, 8010 Graz, AustriaDiagnostic and Research Center for Molecular Biomedicine, Diagnostic and Research Institute of Pathology, Medical University Graz, Neue Stiftingtalstraße 6, 8010 Graz, AustriaDiagnostic and Research Center for Molecular Biomedicine, Diagnostic and Research Institute of Pathology, Medical University Graz, Neue Stiftingtalstraße 6, 8010 Graz, AustriaPivaris BioScience GmbH, Media Quarter Marx 3.4, Maria-Jacobi-Gasse 1, 1030 Vienna, AustriaThe peptide TAT-I24, a fusion of the TAT peptide (amino acids 48–60) and the 9-mer peptide I24, has been previously shown to neutralize several double-stranded (ds) DNA viruses in vitro. We have now extended the testing to potentially sensitive RNA viruses and analyzed the antiviral effect of the peptide against Severe Acute Respiratory Syndrome Coronavirus-2 (SARS-CoV-2). In Vero E6 cells, TAT-I24 neutralized the human 2019-nCoV isolate (Wuhan variant) in a dose-dependent manner, while it was unable to neutralize two SARS-CoV-2 variants of concern, Delta and Omicron. Moreover, TAT-I24 could not significantly neutralize any of the SARS-CoV-2 variants in the human lung carcinoma cell line Calu-3, which provides an alternative entry route for SARS-CoV-2 by direct membrane fusion. Therefore, a possible dependence on virus uptake by endocytosis was investigated by exposing Vero E6 cells to chloroquine (CQ), an inhibitor of endosomal acidification. The Wuhan variant was highly sensitive to inhibition by CQ, an effect which was further enhanced by TAT-I24, while the Delta variant was less sensitive to inhibition by higher concentrations of CQ compared to the Wuhan variant. The microscopic analysis of COS-7 cells using a rhodamine-labeled TAT-I24 (Rho-TAT-I24) showed the endosomal localization of fluorescent TAT-I24 and co-localization with transfected GFP-Rab14 but not GFP-Rab5. As these proteins are found in distinct endosomal pathways, our results indicate that the virus entry pathway determines sensitivity to the peptide.https://www.mdpi.com/1999-4915/17/4/458antiviral peptideSARS-CoV-2virus entryendocytosisRab14
spellingShingle Eva Kicker
Antonio Kouros
Kurt Zatloukal
Hanna Harant
The Virus Entry Pathway Determines Sensitivity to the Antiviral Peptide TAT-I24
Viruses
antiviral peptide
SARS-CoV-2
virus entry
endocytosis
Rab14
title The Virus Entry Pathway Determines Sensitivity to the Antiviral Peptide TAT-I24
title_full The Virus Entry Pathway Determines Sensitivity to the Antiviral Peptide TAT-I24
title_fullStr The Virus Entry Pathway Determines Sensitivity to the Antiviral Peptide TAT-I24
title_full_unstemmed The Virus Entry Pathway Determines Sensitivity to the Antiviral Peptide TAT-I24
title_short The Virus Entry Pathway Determines Sensitivity to the Antiviral Peptide TAT-I24
title_sort virus entry pathway determines sensitivity to the antiviral peptide tat i24
topic antiviral peptide
SARS-CoV-2
virus entry
endocytosis
Rab14
url https://www.mdpi.com/1999-4915/17/4/458
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