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Genome-wide CRISPR knockout screen identifies Activating Transcription Factor (ATF1) as an activator of HIV gene expression that restricts entry into latency

ABSTRACT: Introduction of antiretroviral therapy has limited the spread of HIV and improved clinical outcomes. Yet, a complete cure for infection remains out of reach, as in the face of therapy, a transcriptionally silent but replication-competent provirus persists in a long-lived cell reservoir. Towards identifying new host proteins that regulate HIV gene transcription and latency, we performed a genome-wide CRISPR knockout screen in human CD4+ T cells. We identified the Activating Transcription Factor as a regulator of HIV gene transcription. ATF1 is recruited to the HIV promoter and activates viral gene expression. Gain and loss of function experiments demonstrate that depletion of ATF1 expression decreases HIV gene expression and promotes latency in primary CD4+ infected T cells. Interestingly, ATF1 regulates CCR5 expression and indirectly affects R5-tropic HIV infection. Mechanistically, ATF1 depletion impairs occupancy of RNA Polymerase II on the HIV promoter, and this is accompanied with elevated levels of H3K9me3 repression histone mark around the viral promoter. Genome wide analysis of ATF1 shows that it occupies cellular gene promoters including HIV, thereby controls gene targets that indirectly affect HIV infection. We conclude that ATF1 is an activator of gene transcription that dictates HIV gene expression via both direct and indirect mechanisms.

ORGANISM(S): Homo sapiens

PROVIDER: GSE286022 | GEO | 2025/07/02

REPOSITORIES: GEO

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Project description:Aguilera 2014 - HIV latency. Interaction between HIV proteins and immune response This model is described in the article: Studying HIV latency by modeling the interaction between HIV proteins and the innate immune response. Aguilera LU, Rodríguez-González J. J. Theor. Biol. 2014 Nov; 360: 67-77 Abstract: HIV infection leads to two cell fates, the viral productive state or viral latency (a reversible non-productive state). HIV latency is relevant because infected active CD4+ T-lymphocytes can reach a resting memory state in which the provirus remains silent for long periods of time. Despite experimental and theoretical efforts, the causal molecular mechanisms responsible for HIV latency are only partially understood. Studies have determined that HIV latency is influenced by the innate immune response carried out by cell restriction factors that inhibit the postintegration steps in the virus replication cycle. In this study, we present a mathematical study that combines deterministic and stochastic approaches to analyze the interactions between HIV proteins and the innate immune response. Using wide ranges of parameter values, we observed the following: (1) a phenomenological description of the viral productive and latent cell phenotypes is obtained by bistable and bimodal dynamics, (2) biochemical noise reduces the probability that an infected cell adopts the latent state, (3) the effects of the innate immune response enhance the HIV latency state, (4) the conditions of the cell before infection affect the latent phenotype, i.e., the existing expression of cell restriction factors propitiates HIV latency, and existing expression of HIV proteins reduces HIV latency. This model is hosted on BioModels Database and identified by: BIOMD0000000573. To cite BioModels Database, please use: BioModels Database: An enhanced, curated and annotated resource for published quantitative kinetic models. To the extent possible under law, all copyright and related or neighbouring rights to this encoded model have been dedicated to the public domain worldwide. Please refer to CC0 Public Domain Dedication for more information.

Project description:BACKGROUND: Combination antiretroviral therapy (cART) is able to control HIV-1 viral replication, however long-lived latent infection in resting memory CD4+ T-cells persist. The mechanisms for establishment and maintenance of latent infection in resting memory CD4+ T-cells remain unclear. Previously we have shown that HIV-1 infection of resting CD4+ T-cells co-cultured with CD11c+ myeloid dendritic cells (mDC) produced a population of non-proliferating T-cells with latent infection. Here we asked whether different antigen presenting cells (APC), including subpopulations of DC and monocytes, were able to induce post-integration latent infection in resting CD4+ T-cells, and examined potential cell interactions that may be involved using RNA-seq. RESULTS: mDC (CD1c+), SLAN+ DC and CD14+ monocytes were most efficient in stimulating proliferation of CD4+ T-cells during syngeneic culture and in generating post-integration latent infection in non-proliferating CD4+ T-cells following HIV-1 infection of APC-T-cell co-cultures. In comparison, plasmacytoid DC (pDC) and B-cells did not induce latent infection in APC-T-cell co-cultures. We compared the RNA expression profiles of APC subpopulations that could and could not induce latency in non-proliferating CD4+ T-cells. Gene expression analysis, comparing the mDC, SLAN+ DC and CD14+ monocyte subpopulations to pDC identified 53 upregulated genes that encode proteins expressed on the plasma membrane that could signal to CD4+ T-cells via cell-cell interactions (32 genes), immune checkpoints (IC) (5 genes), T-cell activation (9 genes), regulation of apoptosis (5 genes), antigen presentation (1 gene) and through unknown ligands (1 gene). CONCLUSIONS: APC subpopulations from the myeloid lineage, specifically mDC subpopulations and CD14+ monocytes, were able to efficiently induce post-integration HIV-1 latency in non-proliferating CD4+ T-cells in vitro. Inhibition of key pathways involved in mDC-T-cell interactions and HIV-1 latency may provide novel targets to eliminate HIV latency. mRNA profiles of sorted, pure antigen presenting cells including, CD1c+ myleoid dendirtic cells (mDC), SLAN+ mDC, CD14+ monocytes and plasmacytoid DC (pDC), were generated using next generation sequencing in triplicate, using Illumina Illumina Hiseq 2000.

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Genome-wide CRISPR knockout screen identifies Activating Transcription Factor (ATF1) as an activator of HIV gene expression that restricts entry into latency

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