Pan-retroviral Nucleocapsid-Mediated Phase Separation Regulates Genomic RNA Positioning and Trafficking

Anne Monette; Meijuan Niu; Lois Chen; Shringar Rao; Robert James Gorelick; Andrew John Mouland

doi:10.1016/j.celrep.2020.03.084

Pan-retroviral Nucleocapsid-Mediated Phase Separation Regulates Genomic RNA Positioning and Trafficking

Cell Rep. 2020 Apr 21;31(3):107520. doi: 10.1016/j.celrep.2020.03.084.

Authors

Anne Monette¹, Meijuan Niu², Lois Chen³, Shringar Rao⁴, Robert James Gorelick⁵, Andrew John Mouland⁶

Affiliations

¹ HIV-1 RNA Trafficking Laboratory, Lady Davis Institute at the Jewish General Hospital, Montréal, QC H3T 1E2, Canada. Electronic address: anne.monette@mail.mcgill.ca.
² HIV-1 RNA Trafficking Laboratory, Lady Davis Institute at the Jewish General Hospital, Montréal, QC H3T 1E2, Canada.
³ HIV-1 RNA Trafficking Laboratory, Lady Davis Institute at the Jewish General Hospital, Montréal, QC H3T 1E2, Canada; Department of Microbiology and Immunology, McGill University, Montréal, QC H3A 2B4, Canada.
⁴ HIV-1 RNA Trafficking Laboratory, Lady Davis Institute at the Jewish General Hospital, Montréal, QC H3T 1E2, Canada; Department of Biochemistry, Erasmus University Medical Center, Ee634, PO Box 2040, 3000CA Rotterdam, the Netherlands.
⁵ AIDS and Cancer Virus Program, Frederick National Laboratory for Cancer Research, Frederick, MD 21701, USA.
⁶ HIV-1 RNA Trafficking Laboratory, Lady Davis Institute at the Jewish General Hospital, Montréal, QC H3T 1E2, Canada; Department of Microbiology and Immunology, McGill University, Montréal, QC H3A 2B4, Canada; Department of Medicine, McGill University, Montréal, QC H3G 2M1, Canada. Electronic address: andrew.mouland@mcgill.ca.

Abstract

The duality of liquid-liquid phase separation (LLPS) of cellular components into membraneless organelles defines the nucleation of both normal and disease processes including stress granule (SG) assembly. From mounting evidence of LLPS utility by viruses, we discover that HIV-1 nucleocapsid (NC) protein condenses into zinc-finger (ZnF)-dependent LLPSs that are dynamically influenced by cytosolic factors. ZnF-dependent and Zinc (Zn²⁺)-chelation-sensitive NC-LLPS are formed in live cells. NC-Zn²⁺ ejection reverses the HIV-1 blockade on SG assembly, inhibits NC-SG assembly, disrupts NC/Gag-genomic RNA (vRNA) ribonucleoprotein complexes, and causes nuclear sequestration of NC and the vRNA, inhibiting Gag expression and virus release. NC ZnF mutagenesis eliminates the HIV-1 blockade of SG assembly and repositions vRNA to SGs. We find that NC-mediated, Zn²⁺-coordinated phase separation is conserved among diverse retrovirus subfamilies, illustrating that this exquisitely evolved Zn²⁺-dependent feature of virus replication represents a critical target for pan-antiretroviral therapies.

Keywords: Gag; HIV-1; intrinsically disordered domain; liquid-liquid phase separation; membraneless organelle; nucleocapsid; retrovirus; stress granule; viral genomic RNA trafficking; zinc.

Publication types

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

MeSH terms

Genomics / methods*
Humans
Nucleocapsid / metabolism*
Protein Transport / genetics*
RNA, Viral / genetics*

Substances

RNA, Viral

Abstract

Publication types

MeSH terms

Substances

Grants and funding