tetano
Editor, Senior Moderator
Proc Natl Acad Sci U S A
. 2023 Jan 31;120(5):e2210361120.
doi: 10.1073/pnas.2210361120. Epub 2023 Jan 23.
PCIF1-mediated deposition of 5'-cap N[SUP]6[/SUP],2'- O-dimethyladenosine in ACE2 and TMPRSS2 mRNA regulates susceptibility to SARS-CoV-2 infection
Lingling Wang[SUP] 1 [/SUP], Shaobo Wang[SUP] 1 [/SUP], Lujing Wu[SUP] 1 [/SUP], Wanyu Li[SUP] 1 [/SUP], William Bray[SUP] 1 [/SUP], Alex E Clark[SUP] 2 [/SUP], Gwendolyn Michelle Gonzalez[SUP] 3 4 [/SUP], Yinsheng Wang[SUP] 3 4 [/SUP], Aaron F Carlin[SUP] 2 [/SUP], Tariq M Rana[SUP] 1 [/SUP]
Affiliations
Abstract
Infection with severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) continues to be a major health problem worldwide. Due to the fast emergence of SARS-CoV-2 variants, understanding the molecular mechanisms of viral pathogenesis and developing novel inhibitors are essential and urgent. Here, we investigated the potential roles of N[SUP]6[/SUP],2'-O-dimethyladenosine (m[SUP]6[/SUP]A[SUB]m[/SUB]), one of the most abundant modifications of eukaryotic messenger ribonucleic acid (mRNAs), in SARS-CoV-2 infection of human cells. Using genome-wide m[SUP]6[/SUP]A[SUB]m[/SUB]-exo-seq, RNA sequencing analysis, and Clustered regularly interspaced short palindromic repeats (CRISPR)/Cas9 genome editing, we demonstrate that phosphorylated C-terminal domain (CTD)-interacting factor 1 (PCIF1), a cap-specific adenine N[SUP]6[/SUP]-methyltransferase, plays a major role in facilitating infection of primary human lung epithelial cells and cell lines by SARS-CoV-2, variants of concern, and other coronaviruses. We show that PCIF1 promotes infection by sustaining expression of the coronavirus receptors angiotensin-converting enzyme 2 (ACE2) and transmembrane serine protease 2 (TMPRSS2) via m[SUP]6[/SUP]A[SUB]m[/SUB]-dependent mRNA stabilization. In PCIF1-depleted cells, both ACE2/TMPRSS2 expression and viral infection are rescued by re-expression of wild-type, but not catalytically inactive, PCIF1. These findings suggest a role for PCIF1 and cap m[SUP]6[/SUP]A[SUB]m[/SUB] in regulating SARS-CoV-2 susceptibility and identify a potential therapeutic target for prevention of infection.
Keywords: ACE2; NHBE cells; SARS-CoV-2; TMPRSS2; m6Am methylation.
. 2023 Jan 31;120(5):e2210361120.
doi: 10.1073/pnas.2210361120. Epub 2023 Jan 23.
PCIF1-mediated deposition of 5'-cap N[SUP]6[/SUP],2'- O-dimethyladenosine in ACE2 and TMPRSS2 mRNA regulates susceptibility to SARS-CoV-2 infection
Lingling Wang[SUP] 1 [/SUP], Shaobo Wang[SUP] 1 [/SUP], Lujing Wu[SUP] 1 [/SUP], Wanyu Li[SUP] 1 [/SUP], William Bray[SUP] 1 [/SUP], Alex E Clark[SUP] 2 [/SUP], Gwendolyn Michelle Gonzalez[SUP] 3 4 [/SUP], Yinsheng Wang[SUP] 3 4 [/SUP], Aaron F Carlin[SUP] 2 [/SUP], Tariq M Rana[SUP] 1 [/SUP]
Affiliations
- PMID: 36689652
- DOI: 10.1073/pnas.2210361120
Abstract
Infection with severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) continues to be a major health problem worldwide. Due to the fast emergence of SARS-CoV-2 variants, understanding the molecular mechanisms of viral pathogenesis and developing novel inhibitors are essential and urgent. Here, we investigated the potential roles of N[SUP]6[/SUP],2'-O-dimethyladenosine (m[SUP]6[/SUP]A[SUB]m[/SUB]), one of the most abundant modifications of eukaryotic messenger ribonucleic acid (mRNAs), in SARS-CoV-2 infection of human cells. Using genome-wide m[SUP]6[/SUP]A[SUB]m[/SUB]-exo-seq, RNA sequencing analysis, and Clustered regularly interspaced short palindromic repeats (CRISPR)/Cas9 genome editing, we demonstrate that phosphorylated C-terminal domain (CTD)-interacting factor 1 (PCIF1), a cap-specific adenine N[SUP]6[/SUP]-methyltransferase, plays a major role in facilitating infection of primary human lung epithelial cells and cell lines by SARS-CoV-2, variants of concern, and other coronaviruses. We show that PCIF1 promotes infection by sustaining expression of the coronavirus receptors angiotensin-converting enzyme 2 (ACE2) and transmembrane serine protease 2 (TMPRSS2) via m[SUP]6[/SUP]A[SUB]m[/SUB]-dependent mRNA stabilization. In PCIF1-depleted cells, both ACE2/TMPRSS2 expression and viral infection are rescued by re-expression of wild-type, but not catalytically inactive, PCIF1. These findings suggest a role for PCIF1 and cap m[SUP]6[/SUP]A[SUB]m[/SUB] in regulating SARS-CoV-2 susceptibility and identify a potential therapeutic target for prevention of infection.
Keywords: ACE2; NHBE cells; SARS-CoV-2; TMPRSS2; m6Am methylation.