Journal Article

CRISPR-Cas9 screen of E3 ubiquitin ligases identifies TRAF2 and UHRF1 as regulators of HIV latency in primary human T cells

Bouhaddou, Mehdi, Ott, Melanie, Ochieng' Olwal, Charles, Bediako, Yaw, Eckhardt, Manon, Rathore, Ujjwal, Marson, Alexander, Kaake, Robyn M., Haas, Paige, Hultquist, Judd F., Zuliani-Alvarez, Lorena, Braberg, Hannes, Krogan, Nevan J., Soucheray, Margaret, Easwar Kumar, Vigneshwari, Turner-Groth, Autumn, Hiatt, Joseph, McGregor, Michael J., Haas, Kelsey M., Stevenson, Erica, Swaney, Danielle L.

2024

Overview

HIV, the virus that causes AIDS, heavily relies on the machinery of human cells to infect and replicate. Our study focuses on the host cell’s ubiquitination system which is crucial for numerous cellular processes. Many pathogens, including HIV, exploit this system to enhance their own replication and survival. E3 proteins are part of the ubiquitination pathway that are useful drug targets for host-directed therapies. We interrogated the 116 E3s found in human immune cells known as CD4+ T cells, since these are the target cells infected by HIV. Using CRISPR, a gene-editing tool, we individually removed each of these enzymes and observed the impact on HIV infection in human CD4+ T cells isolated from healthy donors. We discovered that 10 of the E3 enzymes had a significant effect on HIV infection. Two of them, TRAF2 and UHRF1, modulated HIV activity within the cells and triggered an increased release of HIV from previously dormant or “latent” cells in a new primary T cell assay. This finding could guide strategies to perturb hidden HIV reservoirs, a major hurdle to curing HIV. Our study offers insights into HIV-host interactions, identifies new factors that influence HIV infection in immune cells, and introduces a novel methodology for studying HIV infection and latency in human immune cells.

• Publisher: American Society for Microbiology
• Subject: Acquired immune deficiency syndrome / AIDS / Antiviral agents / CCAAT-Enhancer-Binding Proteins - metabolism / CD4 antigen / CD4-Positive T-Lymphocytes / Cells / Chromatography / CRISPR / CRISPR screen / CRISPR-Cas Systems / Enzymes / Epigenetics / Genes / Genomes / Genomics and Proteomics / HIV / HIV - physiology / HIV Infections / HIV latency / Human immunodeficiency virus / Humans / Immune system / Infections / Infectivity / Innate immunity / Latency / Lymphocytes / Lymphocytes T / Mass spectrometry / Proteins / Proteomics / Replication / Research Article / resting primary T cells / Scientific imaging / Therapeutic targets / TNF Receptor-Associated Factor 2 - metabolism / TRAF2 protein / Ubiquitin / ubiquitin E3 ligases / Ubiquitin-Protein Ligases - metabolism / Ubiquitination / Ubiquitins - metabolism / Virus Latency / Virus Replication