TMPRSS11D and TMPRSS13 Activate the SARS-CoV-2 Spike Protein
Abstract
:1. Introduction
2. Materials and Methods
2.1. Cells and Viruses
2.2. Plasmids and Transfection
2.3. Generation of Vero E6 Cells Stably Expressing TTSPs
2.4. Immunoblotting
2.5. Virus Entry Assay
2.6. Multi-Cycle Replication Assay
2.7. Indirect Immunofluorescence Assay (IFA)
2.8. Statistical Analysis
3. Results
3.1. Evaluation of the Effects of TTSPs on the Entry of SARS-CoV-1 and SARS-CoV-2 Using 293T-ACE2 Cells
3.2. TMPRSS11D and 13 Facilitate SARS-CoV-2 Replication
4. Discussion
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Conflicts of Interest
References
- Wu, F.; Zhao, S.; Yu, B.; Chen, Y.-M.; Wang, W.; Song, Z.-G.; Hu, Y.; Tao, Z.-W.; Tian, J.-H.; Pei, Y.-Y.; et al. A New Coronavirus Associated with Human Respiratory Disease in China. Nature 2020, 579, 265–269. [Google Scholar] [CrossRef] [Green Version]
- Zhou, P.; Yang, X.-L.; Wang, X.-G.; Hu, B.; Zhang, L.; Zhang, W.; Si, H.-R.; Zhu, Y.; Li, B.; Huang, C.-L.; et al. A Pneumonia Outbreak Associated with a New Coronavirus of Probable Bat Origin. Nature 2020, 579, 270–273. [Google Scholar] [CrossRef] [Green Version]
- Harrison, A.G.; Lin, T.; Wang, P. Mechanisms of SARS-CoV-2 Transmission and Pathogenesis. Trends Immunol. 2020, 41, 1100–1115. [Google Scholar] [CrossRef] [PubMed]
- Xiao, F.; Tang, M.; Zheng, X.; Liu, Y.; Li, X.; Shan, H. Evidence for Gastrointestinal Infection of SARS-CoV-2. Gastroenterology 2020, 158, 1831–1833.e3. [Google Scholar] [CrossRef] [PubMed]
- Cholankeril, G.; Podboy, A.; Aivaliotis, V.I.; Tarlow, B.; Pham, E.A.; Spencer, S.P.; Kim, D.; Hsing, A.; Ahmed, A. High Prevalence of Concurrent Gastrointestinal Manifestations in Patients With Severe Acute Respiratory Syndrome Coronavirus 2: Early Experience From California. Gastroenterology 2020, 159, 775–777. [Google Scholar] [CrossRef]
- Leung, W.K.; To, K.; Chan, P.K.S.; Chan, H.L.Y.; Wu, A.K.L.; Lee, N.; Yuen, K.Y.; Sung, J.J.Y. Enteric Involvement of Severe Acute Respiratory Syndrome-Associated Coronavirus Infection. Gastroenterology 2003, 125, 1011–1017. [Google Scholar] [CrossRef] [Green Version]
- Huang, J.; Zheng, M.; Tang, X.; Chen, Y.; Tong, A.; Zhou, L. Potential of SARS-CoV-2 to Cause CNS Infection: Biologic Fundamental and Clinical Experience. Front. Neurol. 2020, 11, 659. [Google Scholar] [CrossRef] [PubMed]
- Saleki, K.; Banazadeh, M.; Saghazadeh, A.; Rezaei, N. The Involvement of the Central Nervous System in Patients with COVID-19. Rev. Neurosci. 2020, 31, 453–456. [Google Scholar] [CrossRef]
- Hoffmann, M.; Kleine-Weber, H.; Schroeder, S.; Krüger, N.; Herrler, T.; Erichsen, S.; Schiergens, T.S.; Herrler, G.; Wu, N.-H.; Nitsche, A.; et al. SARS-CoV-2 Cell Entry Depends on ACE2 and TMPRSS2 and Is Blocked by a Clinically Proven Protease Inhibitor. Cell 2020, 181, 271–280.e8. [Google Scholar] [CrossRef] [PubMed]
- Boopathi, S.; Poma, A.B.; Kolandaivel, P. Novel 2019 Coronavirus Structure, Mechanism of Action, Antiviral Drug Promises and Rule out against Its Treatment. J. Biomol. Struct. Dyn. 2020, 1–10. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Cai, Y.; Zhang, J.; Xiao, T.; Peng, H.; Sterling, S.M.; Walsh, R.M., Jr.; Rawson, S.; Rits-Volloch, S.; Chen, B. Distinct Conformational States of SARS-CoV-2 Spike Protein. Science 2020, 369, 1586–1592. [Google Scholar] [CrossRef] [PubMed]
- Tang, T.; Bidon, M.; Jaimes, J.A.; Whittaker, G.R.; Daniel, S. Coronavirus Membrane Fusion Mechanism Offers a Potential Target for Antiviral Development. Antivir. Res. 2020, 178, 104792. [Google Scholar] [CrossRef]
- Hoffmann, M.; Kleine-Weber, H.; Pöhlmann, S. A Multibasic Cleavage Site in the Spike Protein of SARS-CoV-2 Is Essential for Infection of Human Lung Cells. Mol. Cell 2020, 78, 779–784.e5. [Google Scholar] [CrossRef] [PubMed]
- Moreira, R.A.; Guzman, H.V.; Boopathi, S.; Baker, J.L.; Poma, A.B. Characterization of Structural and Energetic Differences between Conformations of the SARS-CoV-2 Spike Protein. Materials 2020, 13, 5362. [Google Scholar] [CrossRef] [PubMed]
- Moreira, R.A.; Chwastyk, M.; Baker, J.L.; Guzman, H.V.; Poma, A.B. Quantitative Determination of Mechanical Stability in the Novel Coronavirus Spike Protein. Nanoscale 2020, 12, 16409–16413. [Google Scholar] [CrossRef]
- Casalino, L.; Gaieb, Z.; Goldsmith, J.A.; Hjorth, C.K.; Dommer, A.C.; Harbison, A.M.; Fogarty, C.A.; Barros, E.P.; Taylor, B.C.; McLellan, J.S.; et al. Beyond Shielding: The Roles of Glycans in the SARS-CoV-2 Spike Protein. ACS Cent. Sci. 2020, 6, 1722–1734. [Google Scholar] [CrossRef]
- Simmons, G.; Zmora, P.; Gierer, S.; Heurich, A.; Pöhlmann, S. Proteolytic Activation of the SARS-Coronavirus Spike Protein: Cutting Enzymes at the Cutting Edge of Antiviral Research. Antivir. Res. 2013, 100, 605–614. [Google Scholar] [CrossRef] [PubMed]
- Ou, T.; Mou, H.; Zhang, L.; Ojha, A.; Choe, H.; Farzan, M. Hydroxychloroquine-Mediated Inhibition of SARS-CoV-2 Entry Is Attenuated by TMPRSS2. PLoS Pathog. 2021, 17, e1009212. [Google Scholar] [CrossRef]
- Matsuyama, S.; Nao, N.; Shirato, K.; Kawase, M.; Saito, S.; Takayama, I.; Nagata, N.; Sekizuka, T.; Katoh, H.; Kato, F.; et al. Enhanced Isolation of SARS-CoV-2 by TMPRSS2-Expressing Cells. Proc. Natl. Acad. Sci. USA 2020, 117, 7001–7003. [Google Scholar] [CrossRef] [Green Version]
- Antalis, T.M.; Bugge, T.H.; Wu, Q. Membrane-Anchored Serine Proteases in Health and Disease. In Progress in Molecular Biology and Translational Science; Elsevier: Amsterdam, The Netherlands, 2011; Volume 99, pp. 1–50. ISBN 978-0-12-385504-6. [Google Scholar]
- Bertram, S.; Glowacka, I.; Blazejewska, P.; Soilleux, E.; Allen, P.; Danisch, S.; Steffen, I.; Choi, S.-Y.; Park, Y.; Schneider, H.; et al. TMPRSS2 and TMPRSS4 Facilitate Trypsin-Independent Spread of Influenza Virus in Caco-2 Cells. J. Virol. 2010, 84, 10016–10025. [Google Scholar] [CrossRef] [Green Version]
- Bertram, S.; Glowacka, I.; Steffen, I.; Kühl, A.; Pöhlmann, S. Novel Insights into Proteolytic Cleavage of Influenza Virus Hemagglutinin: Proteolytic Activation of Influenza Virus. Rev. Med. Virol. 2010, 20, 298–310. [Google Scholar] [CrossRef]
- Laporte, M.; Stevaert, A.; Raeymaekers, V.; Boogaerts, T.; Nehlmeier, I.; Chiu, W.; Benkheil, M.; Vanaudenaerde, B.; Pöhlmann, S.; Naesens, L. Hemagglutinin Cleavability, Acid Stability, and Temperature Dependence Optimize Influenza B Virus for Replication in Human Airways. J. Virol. 2019, 94, e01430-19. [Google Scholar] [CrossRef] [Green Version]
- Shi, W.; Fan, W.; Bai, J.; Tang, Y.; Wang, L.; Jiang, Y.; Tang, L.; Liu, M.; Cui, W.; Xu, Y.; et al. TMPRSS2 and MSPL Facilitate Trypsin-Independent Porcine Epidemic Diarrhea Virus Replication in Vero Cells. Viruses 2017, 9, 114. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Zmora, P.; Hoffmann, M.; Kollmus, H.; Moldenhauer, A.-S.; Danov, O.; Braun, A.; Winkler, M.; Schughart, K.; Pöhlmann, S. TMPRSS11A Activates the Influenza A Virus Hemagglutinin and the MERS Coronavirus Spike Protein and Is Insensitive against Blockade by HAI-1. J. Biol. Chem. 2018, 293, 13863–13873. [Google Scholar] [CrossRef] [Green Version]
- Bertram, S.; Glowacka, I.; Muller, M.A.; Lavender, H.; Gnirss, K.; Nehlmeier, I.; Niemeyer, D.; He, Y.; Simmons, G.; Drosten, C.; et al. Cleavage and Activation of the Severe Acute Respiratory Syndrome Coronavirus Spike Protein by Human Airway Trypsin-Like Protease. J. Virol. 2011, 85, 13363–13372. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Zmora, P.; Blazejewska, P.; Moldenhauer, A.-S.; Welsch, K.; Nehlmeier, I.; Wu, Q.; Schneider, H.; Pohlmann, S.; Bertram, S. DESC1 and MSPL Activate Influenza A Viruses and Emerging Coronaviruses for Host Cell Entry. J. Virol. 2014, 88, 12087–12097. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Xia, S.; Lan, Q.; Su, S.; Wang, X.; Xu, W.; Liu, Z.; Zhu, Y.; Wang, Q.; Lu, L.; Jiang, S. The Role of Furin Cleavage Site in SARS-CoV-2 Spike Protein-Mediated Membrane Fusion in the Presence or Absence of Trypsin. Signal Transduct. Target. Ther. 2020, 5, 92. [Google Scholar] [CrossRef]
- Ou, X.; Liu, Y.; Lei, X.; Li, P.; Mi, D.; Ren, L.; Guo, L.; Guo, R.; Chen, T.; Hu, J.; et al. Characterization of Spike Glycoprotein of SARS-CoV-2 on Virus Entry and Its Immune Cross-Reactivity with SARS-CoV. Nat. Commun. 2020, 11, 1620. [Google Scholar] [CrossRef] [Green Version]
- Zang, R.; Castro, M.F.G.; McCune, B.T.; Zeng, Q.; Rothlauf, P.W.; Sonnek, N.M.; Liu, Z.; Brulois, K.F.; Wang, X.; Greenberg, H.B.; et al. TMPRSS2 and TMPRSS4 Promote SARS-CoV-2 Infection of Human Small Intestinal Enterocytes. Sci. Immunol. 2020, 5, eabc3582. [Google Scholar] [CrossRef] [PubMed]
- Sasaki, M.; Uemura, K.; Sato, A.; Toba, S.; Sanaki, T.; Maenaka, K.; Hall, W.W.; Orba, Y.; Sawa, H. SARS-CoV-2 Variants with Mutations at the S1/S2 Cleavage Site Are Generated in Vitro during Propagation in TMPRSS2-Deficient Cells. PLoS Pathog. 2021, 17, e1009233. [Google Scholar] [CrossRef] [PubMed]
- Thai, H.T.C.; Le, M.Q.; Vuong, C.D.; Parida, M.; Minekawa, H.; Notomi, T.; Hasebe, F.; Morita, K. Development and Evaluation of a Novel Loop-Mediated Isothermal Amplification Method for Rapid Detection of Severe Acute Respiratory Syndrome Coronavirus. J. Clin. Microbiol. 2004, 42, 1956–1961. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Emery, S.L.; Erdman, D.D.; Bowen, M.D.; Newton, B.R.; Winchell, J.M.; Meyer, R.F.; Tong, S.; Cook, B.T.; Holloway, B.P.; McCaustland, K.A.; et al. Real-Time Reverse Transcription–Polymerase Chain Reaction Assay for SARS-Associated Coronavirus. Emerg. Infect. Dis. 2004, 10, 311–316. [Google Scholar] [CrossRef] [PubMed]
- Corman, V.M.; Landt, O.; Kaiser, M.; Molenkamp, R.; Meijer, A.; Chu, D.K.; Bleicker, T.; Brünink, S.; Schneider, J.; Schmidt, M.L.; et al. Detection of 2019 Novel Coronavirus (2019-NCoV) by Real-Time RT-PCR. Eurosurveillance 2020, 25, 2000045. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Overbergh, L.; Kyama, C.M.; Valckx, D.; Debrock, S.; Mwenda, J.M.; Mathieu, C.; D’Hooghe, T. Validation of Real-Time RT-PCR Assays for MRNA Quantification in Baboons. Cytokine 2005, 31, 454–458. [Google Scholar] [CrossRef] [PubMed]
- Matsuyama, S.; Nagata, N.; Shirato, K.; Kawase, M.; Takeda, M.; Taguchi, F. Efficient Activation of the Severe Acute Respiratory Syndrome Coronavirus Spike Protein by the Transmembrane Protease TMPRSS2. J. Virol. 2010, 84, 12658–12664. [Google Scholar] [CrossRef] [Green Version]
- Hörnich, B.F.; Großkopf, A.K.; Schlagowski, S.; Tenbusch, M.; Kleine-Weber, H.; Neipel, F.; Stahl-Hennig, C.; Hahn, A.S. SARS-CoV-2 and SARS-CoV Spike-Mediated Cell-Cell Fusion Differ in the Requirements for Receptor Expression and Proteolytic Activation. J. Virol. 2021, (in press). [Google Scholar] [CrossRef]
- Belouzard, S.; Millet, J.K.; Licitra, B.N.; Whittaker, G.R. Mechanisms of Coronavirus Cell Entry Mediated by the Viral Spike Protein. Viruses 2012, 4, 1011–1033. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Fuentes-Prior, P. Priming of SARS-CoV-2 S Protein by Several Membrane-Bound Serine Proteinases Could Explain Enhanced Viral Infectivity and Systemic COVID-19 Infection. J. Biol. Chem. 2020, 296, 100135. [Google Scholar] [CrossRef] [PubMed]
- Kido, H.; Okumura, Y. MSPL/TMPRSS13. Front. Biosci. 2008, 13, 754–758. [Google Scholar] [CrossRef]
Publisher’s Note: MDPI stays neutral with regard to jurisdictional claims in published maps and institutional affiliations. |
© 2021 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).
Share and Cite
Kishimoto, M.; Uemura, K.; Sanaki, T.; Sato, A.; Hall, W.W.; Kariwa, H.; Orba, Y.; Sawa, H.; Sasaki, M. TMPRSS11D and TMPRSS13 Activate the SARS-CoV-2 Spike Protein. Viruses 2021, 13, 384. https://doi.org/10.3390/v13030384
Kishimoto M, Uemura K, Sanaki T, Sato A, Hall WW, Kariwa H, Orba Y, Sawa H, Sasaki M. TMPRSS11D and TMPRSS13 Activate the SARS-CoV-2 Spike Protein. Viruses. 2021; 13(3):384. https://doi.org/10.3390/v13030384
Chicago/Turabian StyleKishimoto, Mai, Kentaro Uemura, Takao Sanaki, Akihiko Sato, William W. Hall, Hiroaki Kariwa, Yasuko Orba, Hirofumi Sawa, and Michihito Sasaki. 2021. "TMPRSS11D and TMPRSS13 Activate the SARS-CoV-2 Spike Protein" Viruses 13, no. 3: 384. https://doi.org/10.3390/v13030384