Influence of CCL2-mediated modulation of ALIX in the budding and replication of viruses from multiple families.

ABSTRACT: Signaling by C-C motif ligand 2 (CCL2), a β-chemokine, modulates HIV-1 budding and release by mobilizing ALG-2-interacting protein X (ALIX) from the F-actin cytoskeleton to the cytosol. Immunodepleting CCL2 in the medium sequesters ALIX to F-actin. We developed a novel tool to study HIV budding and release without mutating viral late domains or silencing ESCRT genes, but by blocking CCL2 signaling using CRISPR-Cas9 knockout (KO) of the CCL2 or CCR2 genes. We knocked out CCL2 (CCL2KO) and CCR2 (CCR2KO) singly or together (double knockout) in HeLa cells and confirmed that knockout was associated with the absence of CCL2 or CCR2 expression. In KO cells, ALIX was associated with the F-actin cytoskeleton, while in control cells, it was associated with the cytosolic soluble fraction. In KO cells

SUBMITTER: Mofed D 

PROVIDER: S-EPMC12607790 | biostudies-literature | 2025 Nov

REPOSITORIES: biostudies-literature