Effects of CDC5L knockdown on gene transcription levels in HeLa cell lines

Stress granules (SGs) are membraneless condensates that assemble in response to cellular stress. We identified cell division cycle 5-like protein (CDC5L) as a regulator of SG dynamics, where CDC5L knockdown triggered SG formation even in the absence of exogenous stress. Reduced CDC5L levels led to upregulation of PERK at both mRNA and protein levels, suggesting transcriptional regulation. To examine the global impact of CDC5L on gene expression, particularly stress-related pathways, we performed RNA-seq in HeLa cells following siRNA-mediated CDC5L knockdown. In parallel, ChIP-PCR demonstrated that CDC5L directly binds to the PERK promoter, indicating a role as a transcriptional repressor. To further characterize CDC5L-bound genomic regions, we established a stable HeLa cell line expressing CDC5L-HA via lentiviral transduction and performed ChIP-seq to identify promoter regions directly occupied by CDC5L. Together, these transcriptomic and epigenomic datasets define CDC5L as a transcriptional regulator of stress-responsive genes.