Identification of Host Cellular Protein Substrates of SARS-COV-2 Main Protease
The novel severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is the causative agent of coronavirus disease-19 (COVID-19) being associated with severe pneumonia. Like with other viruses, the interaction of SARS-CoV-2 with host cell proteins is necessary for successful replication, and cleav...
| Main Authors: | , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
MDPI AG
2020-12-01
|
| Series: | International Journal of Molecular Sciences |
| Subjects: | |
| Online Access: | https://www.mdpi.com/1422-0067/21/24/9523 |
| _version_ | 1797544655958573056 |
|---|---|
| author | Márió Miczi Mária Golda Balázs Kunkli Tibor Nagy József Tőzsér János András Mótyán |
| author_facet | Márió Miczi Mária Golda Balázs Kunkli Tibor Nagy József Tőzsér János András Mótyán |
| author_sort | Márió Miczi |
| collection | DOAJ |
| description | The novel severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is the causative agent of coronavirus disease-19 (COVID-19) being associated with severe pneumonia. Like with other viruses, the interaction of SARS-CoV-2 with host cell proteins is necessary for successful replication, and cleavage of cellular targets by the viral protease also may contribute to the pathogenesis, but knowledge about the human proteins that are processed by the main protease (3CLpro) of SARS-CoV-2 is still limited. We tested the prediction potentials of two different in silico methods for the identification of SARS-CoV-2 3CLpro cleavage sites in human proteins. Short stretches of homologous host-pathogen protein sequences (SSHHPS) that are present in SARS-CoV-2 polyprotein and human proteins were identified using BLAST analysis, and the NetCorona 1.0 webserver was used to successfully predict cleavage sites, although this method was primarily developed for SARS-CoV. Human C-terminal-binding protein 1 (CTBP1) was found to be cleaved in vitro by SARS-CoV-2 3CLpro, the existence of the cleavage site was proved experimentally by using a His<sub>6</sub>-MBP-mEYFP recombinant substrate containing the predicted target sequence. Our results highlight both potentials and limitations of the tested algorithms. The identification of candidate host substrates of 3CLpro may help better develop an understanding of the molecular mechanisms behind the replication and pathogenesis of SARS-CoV-2. |
| first_indexed | 2024-03-10T14:03:36Z |
| format | Article |
| id | doaj.art-0f1ebdc403ea4d97a20f412e39712e0f |
| institution | Directory Open Access Journal |
| issn | 1661-6596 1422-0067 |
| language | English |
| last_indexed | 2024-03-10T14:03:36Z |
| publishDate | 2020-12-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | International Journal of Molecular Sciences |
| spelling | doaj.art-0f1ebdc403ea4d97a20f412e39712e0f2023-11-21T00:49:30ZengMDPI AGInternational Journal of Molecular Sciences1661-65961422-00672020-12-012124952310.3390/ijms21249523Identification of Host Cellular Protein Substrates of SARS-COV-2 Main ProteaseMárió Miczi0Mária Golda1Balázs Kunkli2Tibor Nagy3József Tőzsér4János András Mótyán5Department of Biochemistry and Molecular Biology, Faculty of Medicine, University of Debrecen, 4032 Debrecen, HungaryDepartment of Biochemistry and Molecular Biology, Faculty of Medicine, University of Debrecen, 4032 Debrecen, HungaryDepartment of Biochemistry and Molecular Biology, Faculty of Medicine, University of Debrecen, 4032 Debrecen, HungaryDepartment of Applied Chemistry, Faculty of Science and Technology, University of Debrecen, 4032 Debrecen, HungaryDepartment of Biochemistry and Molecular Biology, Faculty of Medicine, University of Debrecen, 4032 Debrecen, HungaryDepartment of Biochemistry and Molecular Biology, Faculty of Medicine, University of Debrecen, 4032 Debrecen, HungaryThe novel severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is the causative agent of coronavirus disease-19 (COVID-19) being associated with severe pneumonia. Like with other viruses, the interaction of SARS-CoV-2 with host cell proteins is necessary for successful replication, and cleavage of cellular targets by the viral protease also may contribute to the pathogenesis, but knowledge about the human proteins that are processed by the main protease (3CLpro) of SARS-CoV-2 is still limited. We tested the prediction potentials of two different in silico methods for the identification of SARS-CoV-2 3CLpro cleavage sites in human proteins. Short stretches of homologous host-pathogen protein sequences (SSHHPS) that are present in SARS-CoV-2 polyprotein and human proteins were identified using BLAST analysis, and the NetCorona 1.0 webserver was used to successfully predict cleavage sites, although this method was primarily developed for SARS-CoV. Human C-terminal-binding protein 1 (CTBP1) was found to be cleaved in vitro by SARS-CoV-2 3CLpro, the existence of the cleavage site was proved experimentally by using a His<sub>6</sub>-MBP-mEYFP recombinant substrate containing the predicted target sequence. Our results highlight both potentials and limitations of the tested algorithms. The identification of candidate host substrates of 3CLpro may help better develop an understanding of the molecular mechanisms behind the replication and pathogenesis of SARS-CoV-2.https://www.mdpi.com/1422-0067/21/24/9523COVID-19coronavirusSARSSARS-CoV-2main protease3CL protease |
| spellingShingle | Márió Miczi Mária Golda Balázs Kunkli Tibor Nagy József Tőzsér János András Mótyán Identification of Host Cellular Protein Substrates of SARS-COV-2 Main Protease International Journal of Molecular Sciences COVID-19 coronavirus SARS SARS-CoV-2 main protease 3CL protease |
| title | Identification of Host Cellular Protein Substrates of SARS-COV-2 Main Protease |
| title_full | Identification of Host Cellular Protein Substrates of SARS-COV-2 Main Protease |
| title_fullStr | Identification of Host Cellular Protein Substrates of SARS-COV-2 Main Protease |
| title_full_unstemmed | Identification of Host Cellular Protein Substrates of SARS-COV-2 Main Protease |
| title_short | Identification of Host Cellular Protein Substrates of SARS-COV-2 Main Protease |
| title_sort | identification of host cellular protein substrates of sars cov 2 main protease |
| topic | COVID-19 coronavirus SARS SARS-CoV-2 main protease 3CL protease |
| url | https://www.mdpi.com/1422-0067/21/24/9523 |
| work_keys_str_mv | AT mariomiczi identificationofhostcellularproteinsubstratesofsarscov2mainprotease AT mariagolda identificationofhostcellularproteinsubstratesofsarscov2mainprotease AT balazskunkli identificationofhostcellularproteinsubstratesofsarscov2mainprotease AT tibornagy identificationofhostcellularproteinsubstratesofsarscov2mainprotease AT jozseftozser identificationofhostcellularproteinsubstratesofsarscov2mainprotease AT janosandrasmotyan identificationofhostcellularproteinsubstratesofsarscov2mainprotease |