tetano
Editor, Senior Moderator
Proc Natl Acad Sci U S A
. 2021 Apr 27;118(17):e2023157118.
doi: 10.1073/pnas.2023157118.
ORF10-Cullin-2-ZYG11B complex is not required for SARS-CoV-2 infection
Elijah L Mena[SUP] 1 2 [/SUP], Callie J Donahue[SUP] 3 [/SUP], Laura Pontano Vaites[SUP] 4 [/SUP], Jie Li[SUP] 5 [/SUP], Gergely Rona[SUP] 5 6 [/SUP], Colin O'Leary[SUP] 1 [/SUP], Luca Lignitto[SUP] 5 [/SUP], Bearach Miwatani-Minter[SUP] 5 [/SUP], Joao A Paulo[SUP] 4 [/SUP], Avantika Dhabaria[SUP] 5 7 [/SUP], Beatrix Ueberheide[SUP] 5 7 8 [/SUP], Steven P Gygi[SUP] 4 [/SUP], Michele Pagano[SUP] 9 6 [/SUP], J Wade Harper[SUP] 10 [/SUP], Robert A Davey[SUP] 11 [/SUP], Stephen J Elledge[SUP] 12 2 [/SUP]
Affiliations
Abstract
In order to understand the transmission and virulence of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), it is necessary to understand the functions of each of the gene products encoded in the viral genome. One feature of the SARS-CoV-2 genome that is not present in related, common coronaviruses is ORF10, a putative 38-amino acid protein-coding gene. Proteomic studies found that ORF10 binds to an E3 ubiquitin ligase containing Cullin-2, Rbx1, Elongin B, Elongin C, and ZYG11B (CRL2[SUP]ZYG11B[/SUP]). Since CRL2[SUP]ZYG11B[/SUP] mediates protein degradation, one possible role for ORF10 is to "hijack" CRL2[SUP]ZYG11B[/SUP] in order to target cellular, antiviral proteins for ubiquitylation and subsequent proteasomal degradation. Here, we investigated whether ORF10 hijacks CRL2[SUP]ZYG11B[/SUP] or functions in other ways, for example, as an inhibitor or substrate of CRL2[SUP]ZYG11B[/SUP] While we confirm the ORF10-ZYG11B interaction and show that the N terminus of ORF10 is critical for it, we find no evidence that ORF10 is functioning to inhibit or hijack CRL2[SUP]ZYG11B[/SUP] Furthermore, ZYG11B and its paralog ZER1 are dispensable for SARS-CoV-2 infection in cultured cells. We conclude that the interaction between ORF10 and CRL2[SUP]ZYG11B[/SUP] is not relevant for SARS-CoV-2 infection in vitro.
Keywords: CUL2; ORF10; SARS-CoV-2; ZER1; ZYG11B.
. 2021 Apr 27;118(17):e2023157118.
doi: 10.1073/pnas.2023157118.
ORF10-Cullin-2-ZYG11B complex is not required for SARS-CoV-2 infection
Elijah L Mena[SUP] 1 2 [/SUP], Callie J Donahue[SUP] 3 [/SUP], Laura Pontano Vaites[SUP] 4 [/SUP], Jie Li[SUP] 5 [/SUP], Gergely Rona[SUP] 5 6 [/SUP], Colin O'Leary[SUP] 1 [/SUP], Luca Lignitto[SUP] 5 [/SUP], Bearach Miwatani-Minter[SUP] 5 [/SUP], Joao A Paulo[SUP] 4 [/SUP], Avantika Dhabaria[SUP] 5 7 [/SUP], Beatrix Ueberheide[SUP] 5 7 8 [/SUP], Steven P Gygi[SUP] 4 [/SUP], Michele Pagano[SUP] 9 6 [/SUP], J Wade Harper[SUP] 10 [/SUP], Robert A Davey[SUP] 11 [/SUP], Stephen J Elledge[SUP] 12 2 [/SUP]
Affiliations
- PMID: 33827988
- DOI: 10.1073/pnas.2023157118
Abstract
In order to understand the transmission and virulence of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), it is necessary to understand the functions of each of the gene products encoded in the viral genome. One feature of the SARS-CoV-2 genome that is not present in related, common coronaviruses is ORF10, a putative 38-amino acid protein-coding gene. Proteomic studies found that ORF10 binds to an E3 ubiquitin ligase containing Cullin-2, Rbx1, Elongin B, Elongin C, and ZYG11B (CRL2[SUP]ZYG11B[/SUP]). Since CRL2[SUP]ZYG11B[/SUP] mediates protein degradation, one possible role for ORF10 is to "hijack" CRL2[SUP]ZYG11B[/SUP] in order to target cellular, antiviral proteins for ubiquitylation and subsequent proteasomal degradation. Here, we investigated whether ORF10 hijacks CRL2[SUP]ZYG11B[/SUP] or functions in other ways, for example, as an inhibitor or substrate of CRL2[SUP]ZYG11B[/SUP] While we confirm the ORF10-ZYG11B interaction and show that the N terminus of ORF10 is critical for it, we find no evidence that ORF10 is functioning to inhibit or hijack CRL2[SUP]ZYG11B[/SUP] Furthermore, ZYG11B and its paralog ZER1 are dispensable for SARS-CoV-2 infection in cultured cells. We conclude that the interaction between ORF10 and CRL2[SUP]ZYG11B[/SUP] is not relevant for SARS-CoV-2 infection in vitro.
Keywords: CUL2; ORF10; SARS-CoV-2; ZER1; ZYG11B.