ORF10-Cullin-2-ZYG11B complex is not required for SARS-CoV-2 infection

Elijah L Mena; Callie J Donahue; Laura Pontano Vaites; Jie Li; Gergely Rona; Colin O'Leary; Luca Lignitto; Bearach Miwatani-Minter; Joao A Paulo; Avantika Dhabaria; Beatrix Ueberheide; Steven P Gygi; Michele Pagano; J Wade Harper; Robert A Davey; Stephen J Elledge

doi:10.1073/pnas.2023157118

ORF10-Cullin-2-ZYG11B complex is not required for SARS-CoV-2 infection

Proc Natl Acad Sci U S A. 2021 Apr 27;118(17):e2023157118. doi: 10.1073/pnas.2023157118.

Authors

Elijah L Mena^{1

2}, Callie J Donahue³, Laura Pontano Vaites⁴, Jie Li⁵, Gergely Rona^{5

6}, Colin O'Leary¹, Luca Lignitto⁵, Bearach Miwatani-Minter⁵, Joao A Paulo⁴, Avantika Dhabaria^{5

7}, Beatrix Ueberheide^{5

7

8}, Steven P Gygi⁴, Michele Pagano^{9

6}, J Wade Harper¹⁰, Robert A Davey¹¹, Stephen J Elledge^{12

2}

Affiliations

¹ Division of Genetics, Department of Genetics, Brigham and Women's Hospital, Harvard Medical School, Boston, MA 02115.
² Howard Hughes Medical Institute, Brigham and Women's Hospital, Boston, MA 02115.
³ Department of Microbiology, National Emerging Infectious Disease Laboratories, Boston University Medical Campus, Boston, MA 02118.
⁴ Department of Cell Biology, Blavatnik Institute, Harvard Medical School, Boston, MA 02115.
⁵ Department of Biochemistry and Molecular Pharmacology, New York University Grossman School of Medicine, New York, NY 10016.
⁶ Howard Hughes Medical Institute, New York University Grossman School of Medicine, New York, NY 10016.
⁷ Proteomics Laboratory, Division of Advanced Research Technologies, New York University Grossman School of Medicine, New York, NY 10016.
⁸ Department of Neurology, New York University Grossman School of Medicine, New York, NY 10016.
⁹ Department of Biochemistry and Molecular Pharmacology, New York University Grossman School of Medicine, New York, NY 10016; Michele.Pagano@nyulangone.org wade_harper@hms.harvard.edu radavey@bu.edu selledge@genetics.med.harvard.edu.
¹⁰ Department of Cell Biology, Blavatnik Institute, Harvard Medical School, Boston, MA 02115; Michele.Pagano@nyulangone.org wade_harper@hms.harvard.edu radavey@bu.edu selledge@genetics.med.harvard.edu.
¹¹ Department of Microbiology, National Emerging Infectious Disease Laboratories, Boston University Medical Campus, Boston, MA 02118; Michele.Pagano@nyulangone.org wade_harper@hms.harvard.edu radavey@bu.edu selledge@genetics.med.harvard.edu.
¹² Division of Genetics, Department of Genetics, Brigham and Women's Hospital, Harvard Medical School, Boston, MA 02115; Michele.Pagano@nyulangone.org wade_harper@hms.harvard.edu radavey@bu.edu selledge@genetics.med.harvard.edu.

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^ZYG11B). Since CRL2^ZYG11B mediates protein degradation, one possible role for ORF10 is to "hijack" CRL2^ZYG11B in order to target cellular, antiviral proteins for ubiquitylation and subsequent proteasomal degradation. Here, we investigated whether ORF10 hijacks CRL2^ZYG11B or functions in other ways, for example, as an inhibitor or substrate of CRL2^ZYG11B 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^ZYG11B 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^ZYG11B is not relevant for SARS-CoV-2 infection in vitro.

Keywords: CUL2; ORF10; SARS-CoV-2; ZER1; ZYG11B.

Publication types

Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't

MeSH terms

COVID-19 / genetics
COVID-19 / metabolism*
Cell Cycle Proteins / genetics
Cell Cycle Proteins / metabolism*
Cullin Proteins / genetics
Cullin Proteins / metabolism*
HEK293 Cells
Humans
Multiprotein Complexes / genetics
Multiprotein Complexes / metabolism*
Open Reading Frames*
SARS-CoV-2 / genetics
SARS-CoV-2 / metabolism*
Viral Proteins / genetics
Viral Proteins / metabolism*

Substances

CUL2 protein, human
Cell Cycle Proteins
Cullin Proteins
Multiprotein Complexes
Viral Proteins
ZYG11A protein, human

Abstract

Publication types

MeSH terms

Substances

Grants and funding