Protein Arginylation Is Regulated during SARS-CoV-2 Infection

Author(s):Show less -

Macedo-da-Silva, Janaina ; Rosa-Fernandes, Livia ; Gomes, Vinicius de Morais ; Santiago, Veronica Feijoli ; Santos, Deivid Martins ; Molnar, Catarina Maria Stanischesk ; Barboza, Bruno Rafael ; de Souza, Edmarcia Elisa ; Marques, Rodolfo Ferreira ; Boscardin, Silvia Beatriz ; Durigon, Edison Luiz ; Marinho, Claudio Romero Farias ; Wrenger, Carsten ; Marie, Suely Kazue Nagahashi ; Palmisano, Giuseppe

• Document type: Journal article
• Source: Viruses-Basel; v. 15, n. 2, p. 25-pg., 2023-02-01.

Abstract

Background: In 2019, the world witnessed the onset of an unprecedented pandemic. By February 2022, the infection by SARS-CoV-2 has already been responsible for the death of more than 5 million people worldwide. Recently, we and other groups discovered that SARS-CoV-2 infection induces ER stress and activation of the unfolded protein response (UPR) pathway. Degradation of misfolded/unfolded proteins is an essential element of proteostasis and occurs mainly in lysosomes or proteasomes. The N-terminal arginylation of proteins is characterized as an inducer of ubiquitination and proteasomal degradation by the N-degron pathway. Results: The role of protein arginylation during SARS-CoV-2 infection was elucidated. Protein arginylation was studied in Vero CCL-81, macrophage-like THP1, and Calu-3 cells infected at different times. A reanalysis of in vivo and in vitro public omics data combined with immunoblotting was performed to measure levels of arginyl-tRNA-protein transferase (ATE1) and its substrates. Dysregulation of the N-degron pathway was specifically identified during coronavirus infections compared to other respiratory viruses. We demonstrated that during SARS-CoV-2 infection, there is an increase in ATE1 expression in Calu-3 and Vero CCL-81 cells. On the other hand, infected macrophages showed no enzyme regulation. ATE1 and protein arginylation was variant-dependent, as shown using P1 and P2 viral variants and HEK 293T cells transfection with the spike protein and receptor-binding domains (RBD). In addition, we report that ATE1 inhibitors, tannic acid and merbromine (MER) reduce viral load. This finding was confirmed in ATE1-silenced cells. Conclusions: We demonstrate that ATE1 is increased during SARS-CoV-2 infection and its inhibition has potential therapeutic value. (AU)

• FAPESP's process: 18/15549-1 - Post-translational modifications in Chagas Disease biological processes and diagnostics: novel methodological approaches and biological applications
• Grantee: Giuseppe Palmisano
• Support Opportunities: Research Grants - Young Investigators Grants - Phase 2
• FAPESP's process: 18/18257-1 - Multi-user equipment approved in grant 14/06863-3: HPLC system configured for analysis of carbohydrates, amino acidis, peptides and glycoproteins
• Grantee: Giuseppe Palmisano
• Support Opportunities: Multi-user Equipment Program
• FAPESP's process: 20/06409-1 - Evaluation of humoral immune response and inflammatory response in patients with confirmed diagnosis of COVID-19 at Hospital Sírio Libanês and correlation with disease severity
• Grantee: Edison Luiz Durigon
• Support Opportunities: Regular Research Grants
• FAPESP's process: 22/09915-0 - Modulation of host N-glycosylation machinery during Trypanosoma cruzi infection
• Grantee: Bruno Rafael Barboza
• Support Opportunities: Scholarships in Brazil - Doctorate
• FAPESP's process: 20/02988-7 - Decoding the impact of microenvironment and signaling pathways in health and disease in brain, adrenal gland and kidney
• Grantee: Suely Kazue Nagahashi Marie
• Support Opportunities: Research Projects - Thematic Grants
• FAPESP's process: 21/00140-3 - Understanding the role of protein arginylation in health and disease through analytical and biological approaches
• Grantee: Janaína Macedo da Silva
• Support Opportunities: Scholarships in Brazil - Doctorate
• FAPESP's process: 20/12277-0 - Drug discovery against human infectious diseases
• Grantee: Edmarcia Elisa de Souza
• Support Opportunities: Scholarships in Brazil - Post-Doctoral
• FAPESP's process: 21/14179-9 - The role of the endoplasmic reticulum in viral and parasitic virulence
• Grantee: Deivid Martins Santos
• Support Opportunities: Scholarships in Brazil - Doctorate (Direct)
• FAPESP's process: 15/26722-8 - Drug discovery against human infectious diseases
• Grantee: Carsten Wrenger
• Support Opportunities: Research Projects - Thematic Grants