tetano
Editor, Senior Moderator
J Biol Chem
. 2020 Jun 22;jbc.AC120.013449.
doi: 10.1074/jbc.AC120.013449. Online ahead of print.
Structural and Functional Conservation of the Programmed -1 Ribosomal Frameshift Signal of SARS Coronavirus 2 (SARS-CoV-2)
Jamie A Kelly[SUP] 1 [/SUP], Alexandra N Olson[SUP] 2 [/SUP], Krishna Neupane[SUP] 3 [/SUP], Sneha Munshi[SUP] 3 [/SUP], Josue San Emeterio[SUP] 4 [/SUP], Lois Pollack[SUP] 5 [/SUP], Michael T Woodside[SUP] 3 [/SUP], Jonathan D Dinman[SUP] 6 [/SUP]
Affiliations
Abstract
About 17 years after the severe acute respiratory syndrome coronavirus (SARS-CoV) epidemic, the world is currently facing the COVID-19 pandemic caused by SARS coronavirus 2 (SARS-CoV-2). According to the most optimistic projections, it will take more than a year to develop a vaccine, so the best short-term strategy may lie in identifying virus-specific targets for small molecule-based interventions. All coronaviruses utilize a molecular mechanism called programmed -1 ribosomal frameshift (-1 PRF) to control the relative expression of their proteins. Previous analyses of SARS-CoV have revealed that it employs a structurally unique three-stemmed mRNA pseudoknot that stimulates high -1 PRF rates and that it also harbors a -1 PRF attenuation element. Altering -1 PRF activity impairs virus replication, suggesting that this activity may be therapeutically targeted. Here, we comparatively analyzed the SARS-CoV and SARS-CoV-2 frameshift signals. Structural and functional analyses revealed that both elements promote similar -1 PRF rates and that silent coding mutations in the slippery sites and in all three stems of the pseudoknot strongly ablate -1 PRF activity. We noted that the upstream attenuator hairpin activity is also functionally retained in both viruses, despite differences in the primary sequence in this region. Small-angle X-ray scattering analyses indicated that the pseudoknots in SARS-CoV and SARS-CoV-2 have the same conformation. Finally, a small molecule previously shown to bind the SARS-CoV pseudoknot and inhibit -1 PRF was similarly effective against -1 PRF in SARS-CoV-2, suggesting that such frameshift inhibitors may be promising lead compounds to combat the current COVID-19 pandemic.
Keywords: (+) ssRNA; RNA; RNA structure; coronavirus; coronavirus disease 2019 (COVID-19); inhibitor; programmed −1 ribosomal frameshifting (−1 PRF); small molecule inhibitor; translation; virus.
. 2020 Jun 22;jbc.AC120.013449.
doi: 10.1074/jbc.AC120.013449. Online ahead of print.
Structural and Functional Conservation of the Programmed -1 Ribosomal Frameshift Signal of SARS Coronavirus 2 (SARS-CoV-2)
Jamie A Kelly[SUP] 1 [/SUP], Alexandra N Olson[SUP] 2 [/SUP], Krishna Neupane[SUP] 3 [/SUP], Sneha Munshi[SUP] 3 [/SUP], Josue San Emeterio[SUP] 4 [/SUP], Lois Pollack[SUP] 5 [/SUP], Michael T Woodside[SUP] 3 [/SUP], Jonathan D Dinman[SUP] 6 [/SUP]
Affiliations
- PMID: 32571880
- DOI: 10.1074/jbc.AC120.013449
Abstract
About 17 years after the severe acute respiratory syndrome coronavirus (SARS-CoV) epidemic, the world is currently facing the COVID-19 pandemic caused by SARS coronavirus 2 (SARS-CoV-2). According to the most optimistic projections, it will take more than a year to develop a vaccine, so the best short-term strategy may lie in identifying virus-specific targets for small molecule-based interventions. All coronaviruses utilize a molecular mechanism called programmed -1 ribosomal frameshift (-1 PRF) to control the relative expression of their proteins. Previous analyses of SARS-CoV have revealed that it employs a structurally unique three-stemmed mRNA pseudoknot that stimulates high -1 PRF rates and that it also harbors a -1 PRF attenuation element. Altering -1 PRF activity impairs virus replication, suggesting that this activity may be therapeutically targeted. Here, we comparatively analyzed the SARS-CoV and SARS-CoV-2 frameshift signals. Structural and functional analyses revealed that both elements promote similar -1 PRF rates and that silent coding mutations in the slippery sites and in all three stems of the pseudoknot strongly ablate -1 PRF activity. We noted that the upstream attenuator hairpin activity is also functionally retained in both viruses, despite differences in the primary sequence in this region. Small-angle X-ray scattering analyses indicated that the pseudoknots in SARS-CoV and SARS-CoV-2 have the same conformation. Finally, a small molecule previously shown to bind the SARS-CoV pseudoknot and inhibit -1 PRF was similarly effective against -1 PRF in SARS-CoV-2, suggesting that such frameshift inhibitors may be promising lead compounds to combat the current COVID-19 pandemic.
Keywords: (+) ssRNA; RNA; RNA structure; coronavirus; coronavirus disease 2019 (COVID-19); inhibitor; programmed −1 ribosomal frameshifting (−1 PRF); small molecule inhibitor; translation; virus.