Proteomic Identification of Potential Target Proteins of Cathepsin W for Its Development as a Drug Target for Influenza
ABSTRACT Influenza A virus (IAV) coopts numerous host factors for efficient replication. The cysteine protease cathepsin W (CTSW) has been identified as one host factor required for IAV entry, specifically for the escape of IAVs from late endosomes. However, the substrate specificity of CTSW and the...
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| Format: | Article |
| Language: | English |
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American Society for Microbiology
2022-08-01
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| Series: | Microbiology Spectrum |
| Subjects: | |
| Online Access: | https://journals.asm.org/doi/10.1128/spectrum.00921-22 |
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| author | Sira C. Günther Carles Martínez-Romero Milagros Sempere Borau Christine T. N. Pham Adolfo García-Sastre Silke Stertz |
| author_facet | Sira C. Günther Carles Martínez-Romero Milagros Sempere Borau Christine T. N. Pham Adolfo García-Sastre Silke Stertz |
| author_sort | Sira C. Günther |
| collection | DOAJ |
| description | ABSTRACT Influenza A virus (IAV) coopts numerous host factors for efficient replication. The cysteine protease cathepsin W (CTSW) has been identified as one host factor required for IAV entry, specifically for the escape of IAVs from late endosomes. However, the substrate specificity of CTSW and the proviral mechanism are thus far unknown. Here, we show that intracellular but not secreted CTSW promotes viral entry. We reveal 79 potential direct and 31 potential indirect cellular target proteins of CTSW using the high-throughput proteomic approach terminal amine isotopic labeling of substrates (TAILS) and determine the cleavage motif shared by the substrates of CTSW. Subsequent integration with data from RNA interference (RNAi) screens for IAV host factors uncovers first insights into the proviral function of CTSW. Notably, CTSW-deficient mice display a 25% increase in survival and a delay in mortality compared to wild-type mice upon IAV infection. Altogether, these findings support the development of drugs targeting CTSW as novel host-directed antiviral therapies. IMPORTANCE Influenza viruses are respiratory pathogens and pose a constant threat to human health. Although antiviral drugs are available for influenza, the emergence and spread of drug-resistant viruses is cause for concern. Therefore, the development of new antivirals with lower chances of their target viruses acquiring resistance is urgently needed to reduce the high morbidity and mortality caused by influenza. Promising alternatives to drugs targeting viral proteins are those directed against host factors required for viral replication. The cysteine protease cathepsin W (CTSW) is an important host factor for IAV replication, and its proteolytic activity is required for fusion of viral and endosomal membranes. In this work, we identify a number of hitherto unknown CTSW substrates, providing new insights into virus-host interactions, and reveal that CTSW might also play a proviral role in an in vivo model. These results support the development of CTSW as a drug target for next-generation antivirals against influenza. |
| first_indexed | 2024-04-13T18:03:23Z |
| format | Article |
| id | doaj.art-328d2796bc78464390cf1a5669260d23 |
| institution | Directory Open Access Journal |
| issn | 2165-0497 |
| language | English |
| last_indexed | 2024-04-13T18:03:23Z |
| publishDate | 2022-08-01 |
| publisher | American Society for Microbiology |
| record_format | Article |
| series | Microbiology Spectrum |
| spelling | doaj.art-328d2796bc78464390cf1a5669260d232022-12-22T02:36:08ZengAmerican Society for MicrobiologyMicrobiology Spectrum2165-04972022-08-0110410.1128/spectrum.00921-22Proteomic Identification of Potential Target Proteins of Cathepsin W for Its Development as a Drug Target for InfluenzaSira C. Günther0Carles Martínez-Romero1Milagros Sempere Borau2Christine T. N. Pham3Adolfo García-Sastre4Silke Stertz5Institute of Medical Virology, University of Zurich, Zurich, SwitzerlandDepartment of Microbiology, Icahn School of Medicine at Mount Sinai, New York, New York, USAInstitute of Medical Virology, University of Zurich, Zurich, SwitzerlandDepartment of Medicine, Division of Rheumatology, Washington University School of Medicine, St. Louis, Missouri, USADepartment of Microbiology, Icahn School of Medicine at Mount Sinai, New York, New York, USAInstitute of Medical Virology, University of Zurich, Zurich, SwitzerlandABSTRACT Influenza A virus (IAV) coopts numerous host factors for efficient replication. The cysteine protease cathepsin W (CTSW) has been identified as one host factor required for IAV entry, specifically for the escape of IAVs from late endosomes. However, the substrate specificity of CTSW and the proviral mechanism are thus far unknown. Here, we show that intracellular but not secreted CTSW promotes viral entry. We reveal 79 potential direct and 31 potential indirect cellular target proteins of CTSW using the high-throughput proteomic approach terminal amine isotopic labeling of substrates (TAILS) and determine the cleavage motif shared by the substrates of CTSW. Subsequent integration with data from RNA interference (RNAi) screens for IAV host factors uncovers first insights into the proviral function of CTSW. Notably, CTSW-deficient mice display a 25% increase in survival and a delay in mortality compared to wild-type mice upon IAV infection. Altogether, these findings support the development of drugs targeting CTSW as novel host-directed antiviral therapies. IMPORTANCE Influenza viruses are respiratory pathogens and pose a constant threat to human health. Although antiviral drugs are available for influenza, the emergence and spread of drug-resistant viruses is cause for concern. Therefore, the development of new antivirals with lower chances of their target viruses acquiring resistance is urgently needed to reduce the high morbidity and mortality caused by influenza. Promising alternatives to drugs targeting viral proteins are those directed against host factors required for viral replication. The cysteine protease cathepsin W (CTSW) is an important host factor for IAV replication, and its proteolytic activity is required for fusion of viral and endosomal membranes. In this work, we identify a number of hitherto unknown CTSW substrates, providing new insights into virus-host interactions, and reveal that CTSW might also play a proviral role in an in vivo model. These results support the development of CTSW as a drug target for next-generation antivirals against influenza.https://journals.asm.org/doi/10.1128/spectrum.00921-22cysteine protease cathepsin WCTSWinfluenza A virusterminal amine isotopic labeling of substratesTAILSepsin 2 |
| spellingShingle | Sira C. Günther Carles Martínez-Romero Milagros Sempere Borau Christine T. N. Pham Adolfo García-Sastre Silke Stertz Proteomic Identification of Potential Target Proteins of Cathepsin W for Its Development as a Drug Target for Influenza Microbiology Spectrum cysteine protease cathepsin W CTSW influenza A virus terminal amine isotopic labeling of substrates TAILS epsin 2 |
| title | Proteomic Identification of Potential Target Proteins of Cathepsin W for Its Development as a Drug Target for Influenza |
| title_full | Proteomic Identification of Potential Target Proteins of Cathepsin W for Its Development as a Drug Target for Influenza |
| title_fullStr | Proteomic Identification of Potential Target Proteins of Cathepsin W for Its Development as a Drug Target for Influenza |
| title_full_unstemmed | Proteomic Identification of Potential Target Proteins of Cathepsin W for Its Development as a Drug Target for Influenza |
| title_short | Proteomic Identification of Potential Target Proteins of Cathepsin W for Its Development as a Drug Target for Influenza |
| title_sort | proteomic identification of potential target proteins of cathepsin w for its development as a drug target for influenza |
| topic | cysteine protease cathepsin W CTSW influenza A virus terminal amine isotopic labeling of substrates TAILS epsin 2 |
| url | https://journals.asm.org/doi/10.1128/spectrum.00921-22 |
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