TY - JOUR
T1 - CRISPR-Cas9 screen of E3 ubiquitin ligases identifies TRAF2 and UHRF1 as regulators of HIV latency in primary human T cells
AU - Rathore, Ujjwal
AU - Haas, Paige
AU - Kumar, Vigneshwari Easwar
AU - Hiatt, Joseph
AU - Haas, Kelsey M.
AU - Bouhaddou, Mehdi
AU - Swaney, Danielle L.
AU - Stevenson, Erica
AU - Zuliani-Alvarez, Lorena
AU - McGregor, Michael J.
AU - Turner-Groth, Autumn
AU - Olwal, Charles Ochieng
AU - Bediako, Yaw
AU - Braberg, Hannes
AU - Soucheray, Margaret
AU - Ott, Melanie
AU - Eckhardt, Manon
AU - Hultquist, Judd F.
AU - Marson, Alexander
AU - Kaake, Robyn M.
AU - Krogan, Nevan J.
N1 - Publisher Copyright:
© 2024 Rathore et al.
PY - 2024/4
Y1 - 2024/4
N2 - During HIV infection of CD4+ T cells, ubiquitin pathways are essential to viral replication and host innate immune response; however, the role of specific E3 ubiquitin ligases is not well understood. Proteomics analyses identified 116 single-subunit E3 ubiquitin ligases expressed in activated primary human CD4+ T cells. Using a CRISPR-based arrayed spreading infectivity assay, we systematically knocked out 116 E3s from activated primary CD4+ T cells and infected them with NL4-3 GFP reporter HIV-1. We found 10 E3s significantly positively or negatively affected HIV infection in activated primary CD4+ T cells, including UHRF1 (pro-viral) and TRAF2 (anti-viral). Furthermore, deletion of either TRAF2 or UHRF1 in three JLat models of latency spontaneously increased HIV transcription. To verify this effect, we developed a CRISPR-compatible resting primary human CD4+ T cell model of latency. Using this system, we found that deletion of TRAF2 or UHRF1 initiated latency reactivation and increased virus production from primary human resting CD4+ T cells, suggesting these two E3s represent promising targets for future HIV latency reversal strategies.
AB - During HIV infection of CD4+ T cells, ubiquitin pathways are essential to viral replication and host innate immune response; however, the role of specific E3 ubiquitin ligases is not well understood. Proteomics analyses identified 116 single-subunit E3 ubiquitin ligases expressed in activated primary human CD4+ T cells. Using a CRISPR-based arrayed spreading infectivity assay, we systematically knocked out 116 E3s from activated primary CD4+ T cells and infected them with NL4-3 GFP reporter HIV-1. We found 10 E3s significantly positively or negatively affected HIV infection in activated primary CD4+ T cells, including UHRF1 (pro-viral) and TRAF2 (anti-viral). Furthermore, deletion of either TRAF2 or UHRF1 in three JLat models of latency spontaneously increased HIV transcription. To verify this effect, we developed a CRISPR-compatible resting primary human CD4+ T cell model of latency. Using this system, we found that deletion of TRAF2 or UHRF1 initiated latency reactivation and increased virus production from primary human resting CD4+ T cells, suggesting these two E3s represent promising targets for future HIV latency reversal strategies.
KW - CRISPR screen
KW - HIV latency
KW - human immunodeficiency virus
KW - resting primary T cells
KW - ubiquitin E3 ligases
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U2 - 10.1128/mbio.02222-23
DO - 10.1128/mbio.02222-23
M3 - Article
C2 - 38411080
AN - SCOPUS:85190482388
SN - 2161-2129
VL - 15
JO - mBio
JF - mBio
IS - 4
ER -