Dataset Information

Systematic discovery of pro- and anti-HIV host factors in primary human CD4+ T cells

ABSTRACT: Host factors that promote or restrict human immunodeficiency virus (HIV) infection in human CD4+ T cells have not been comprehensively identified. We performed orthogonal genome-wide CRISPR activation (CRISPRa) and CRISPR knockout (CRISPRn) screens in primary human CD4+ T cells to systematically discover pro- and anti-HIV host factors. Candidate hits were validated using secondary pooled screens and arrayed perturbations and were further characterized using assays for HIV infection, T cell activation, and HIV receptor/co-receptor expression. CRISPRa identified multiple potent antiviral factors including PI16, PPID, SHISA3, and ITM2A. PI16 interacts with host pathways involved in HIV fusion and inhibits viral entry, while PPID (Cyp40) binds HIV capsid and restricts infection by reducing nuclear import of the HIV core. Structural modeling, evolutionary analyses, and targeted mutagenesis identified domains and residues required for PPID-mediated restriction, including non-human primate ortholog substitutions that enhance antiviral activity. Together, these data define the functional HIV–host interaction landscape in primary human CD4+ T cells and reveal new mechanisms modulating HIV infection.

ORGANISM(S): synthetic construct Homo sapiens

PROVIDER: GSE318876 | GEO | 2026/04/23

REPOSITORIES: GEO

ACCESS DATA

Dataset's files

Source:

			Action	DRS
		Other

Items per page:

1 - 1 of 1

Similar Datasets

Project description:Although HIV-1 can directly infect resting CD4+ T cells, virus replication in resting CD4+T cells is very inefficient owing to the different host restriction factors blocking viral replication. The accessory protein Vpx from the major simian immunodeficiency virus (SIV) of rhesus macaque (mac) and HIV-2 lineage could degrade a host restriction factor, SAM and HD domain containing protein 1 (SAMHD1), to facilitate HIV reverse transcription. Interestingly, Vpx proteins from a second SIV lineage, the SIV of redcapped mangabeys or mandrills (SIVrcm/nmd-2), had no effect on SAMHD1 and did not affect the dNTP pool, but strongly increased HIV-1 infection in resting CD4+ T cells although not in primary macrophages. This indicates that Vpx, in addition to SAMHD1,can overcome a previously unexplored restriction factor for lentiviruses. Here to identify this potential restriction factor, we examined Vpxrcm-interacting cellular proteins and found that keratin 72 (KRT72), an intermediate filament protein that is exclusively expressed in resting CD4+ T cells, is a new host antiviral factor targeted by Vpx. Other than Vpx from SIV mac and HIV-2, the Vpxrcm/nmd-2 lineage, which had no effect on the SAMHD1 protein, could strongly promote the degradation of KRT72, resulting in enhanced HIV-1 infection in resting CD4+ T cells. Furthermore, we discovered that KRT72 restricts HIV-1 replication by sequestering incoming HIV-1 capsids in cytoplasmic intermediate filaments (IFs). In the presence of KRT72, HIV-1 capsid cores become attached to the IF and their trafficking toward the nucleus is inhibited. In contrast, in the absence of KRT72, HIV-1 capsids are transported into the nucleus,leading to high levels of integrated HIV-1 DNA. In addition, KRT72 expression was substantially higher in resting CD4+ T cells than in activated CD4+ T cells, and it was rapidly reduced by T cell activation. Collectively, the results show that KRT72 is a new Vpx-counteracted host antiviral factor that acts to tether incoming capsids to the cytoplasmic IF, thereby restricting HIV-1 infection in resting CD4+ T cells.

Dataset Information

Systematic discovery of pro- and anti-HIV host factors in primary human CD4+ T cells

Dataset's files

Similar Datasets

OmicsDI is part of the ELIXIR infrastructure

Tweets