tetano
Editor, Senior Moderator
Genes Dev
. 2021 Jun 24.
doi: 10.1101/gad.348320.121. Online ahead of print.
Targeting the m [SUP]6[/SUP] A RNA modification pathway blocks SARS-CoV-2 and HCoV-OC43 replication
Hannah M Burgess[SUP] 1 [/SUP], Daniel P Depledge[SUP] 2 [/SUP], Letitia Thompson[SUP] 1 [/SUP], Kalanghad Puthankalam Srinivas[SUP] 1 [/SUP], Rebecca C Grande[SUP] 1 [/SUP], Elizabeth I Vink[SUP] 1 [/SUP], Jonathan S Abebe[SUP] 2 [/SUP], Wesley P Blackaby[SUP] 3 [/SUP], Alan Hendrick[SUP] 3 [/SUP], Mark R Albertella[SUP] 3 [/SUP], Tony Kouzarides[SUP] 4 [/SUP], Kenneth A Stapleford[SUP] 1 [/SUP], Angus C Wilson[SUP] 1 [/SUP], Ian Mohr[SUP] 1 [/SUP]
Affiliations
Abstract
N[SUP]6[/SUP]-methyladenosine (m[SUP]6[/SUP]A) is an abundant internal RNA modification, influencing transcript fate and function in uninfected and virus-infected cells. Installation of m[SUP]6[/SUP]A by the nuclear RNA methyltransferase METTL3 occurs cotranscriptionally; however, the genomes of some cytoplasmic RNA viruses are also m[SUP]6[/SUP]A-modified. How the cellular m[SUP]6[/SUP]A modification machinery impacts coronavirus replication, which occurs exclusively in the cytoplasm, is unknown. Here we show that replication of SARS-CoV-2, the agent responsible for the COVID-19 pandemic, and a seasonal human β-coronavirus HCoV-OC43, can be suppressed by depletion of METTL3 or cytoplasmic m[SUP]6[/SUP]A reader proteins YTHDF1 and YTHDF3 and by a highly specific small molecule METTL3 inhibitor. Reduction of infectious titer correlates with decreased synthesis of viral RNAs and the essential nucleocapsid (N) protein. Sites of m[SUP]6[/SUP]A modification on genomic and subgenomic RNAs of both viruses were mapped by methylated RNA immunoprecipitation sequencing (meRIP-seq). Levels of host factors involved in m[SUP]6[/SUP]A installation, removal, and recognition were unchanged by HCoV-OC43 infection; however, nuclear localization of METTL3 and cytoplasmic m[SUP]6[/SUP]A readers YTHDF1 and YTHDF2 increased. This establishes that coronavirus RNAs are m[SUP]6[/SUP]A-modified and host m[SUP]6[/SUP]A pathway components control β-coronavirus replication. Moreover, it illustrates the therapeutic potential of targeting the m[SUP]6[/SUP]A pathway to restrict coronavirus reproduction.
Keywords: HCoV-OC43; N6-methyladenosine; RNA modification; SARS-CoV-2; coronavirus; direct RNA sequencing; nanopore sequencing; virus–host interactions.
. 2021 Jun 24.
doi: 10.1101/gad.348320.121. Online ahead of print.
Targeting the m [SUP]6[/SUP] A RNA modification pathway blocks SARS-CoV-2 and HCoV-OC43 replication
Hannah M Burgess[SUP] 1 [/SUP], Daniel P Depledge[SUP] 2 [/SUP], Letitia Thompson[SUP] 1 [/SUP], Kalanghad Puthankalam Srinivas[SUP] 1 [/SUP], Rebecca C Grande[SUP] 1 [/SUP], Elizabeth I Vink[SUP] 1 [/SUP], Jonathan S Abebe[SUP] 2 [/SUP], Wesley P Blackaby[SUP] 3 [/SUP], Alan Hendrick[SUP] 3 [/SUP], Mark R Albertella[SUP] 3 [/SUP], Tony Kouzarides[SUP] 4 [/SUP], Kenneth A Stapleford[SUP] 1 [/SUP], Angus C Wilson[SUP] 1 [/SUP], Ian Mohr[SUP] 1 [/SUP]
Affiliations
- PMID: 34168039
- DOI: 10.1101/gad.348320.121
Abstract
N[SUP]6[/SUP]-methyladenosine (m[SUP]6[/SUP]A) is an abundant internal RNA modification, influencing transcript fate and function in uninfected and virus-infected cells. Installation of m[SUP]6[/SUP]A by the nuclear RNA methyltransferase METTL3 occurs cotranscriptionally; however, the genomes of some cytoplasmic RNA viruses are also m[SUP]6[/SUP]A-modified. How the cellular m[SUP]6[/SUP]A modification machinery impacts coronavirus replication, which occurs exclusively in the cytoplasm, is unknown. Here we show that replication of SARS-CoV-2, the agent responsible for the COVID-19 pandemic, and a seasonal human β-coronavirus HCoV-OC43, can be suppressed by depletion of METTL3 or cytoplasmic m[SUP]6[/SUP]A reader proteins YTHDF1 and YTHDF3 and by a highly specific small molecule METTL3 inhibitor. Reduction of infectious titer correlates with decreased synthesis of viral RNAs and the essential nucleocapsid (N) protein. Sites of m[SUP]6[/SUP]A modification on genomic and subgenomic RNAs of both viruses were mapped by methylated RNA immunoprecipitation sequencing (meRIP-seq). Levels of host factors involved in m[SUP]6[/SUP]A installation, removal, and recognition were unchanged by HCoV-OC43 infection; however, nuclear localization of METTL3 and cytoplasmic m[SUP]6[/SUP]A readers YTHDF1 and YTHDF2 increased. This establishes that coronavirus RNAs are m[SUP]6[/SUP]A-modified and host m[SUP]6[/SUP]A pathway components control β-coronavirus replication. Moreover, it illustrates the therapeutic potential of targeting the m[SUP]6[/SUP]A pathway to restrict coronavirus reproduction.
Keywords: HCoV-OC43; N6-methyladenosine; RNA modification; SARS-CoV-2; coronavirus; direct RNA sequencing; nanopore sequencing; virus–host interactions.