G-quadruplex structural dynamics at MAPK12 promoter dictates transcriptional switch to determine stemness in breast cancer.
Pallabi SenguptaAnindya DuttaY V SuseelaTanaya RoychowdhuryNilanjan BanerjeeAnanya DuttaSatyajit HalderKuladip JanaGopeswar MukherjeeSamit ChattopadhyayThimmaiah GovindarajuSubhrangsu ChatterjeePublished in: Cellular and molecular life sciences : CMLS (2024)
P38γ (MAPK12) is predominantly expressed in triple negative breast cancer cells (TNBC) and induces stem cell (CSC) expansion resulting in decreased survival of the patients due to metastasis. Abundance of G-rich sequences at MAPK12 promoter implied the functional probability to reverse tumorigenesis, though the formation of G-Quadruplex (G4) structures at MAPK12 promoter is elusive. Here, we identified two evolutionary consensus adjacent G4 motifs upstream of the MAPK12 promoter, forming parallel G4 structures. They exist in an equilibria between G4 and duplex, regulated by the binding turnover of Sp1 and Nucleolin that bind to these G4 motifs and regulate MAPK12 transcriptional homeostasis. To underscore the gene-regulatory functions of G4 motifs, we employed CRISPR-Cas9 system to eliminate G4s from TNBC cells and synthesized a naphthalene diimide (NDI) derivative (TGS24) which shows high-affinity binding to MAPK12-G4 and inhibits MAPK12 transcription. Deletion of G4 motifs and NDI compound interfere with the recruitment of the transcription factors, inhibiting MAPK12 expression in cancer cells. The molecular basis of NDI-induced G4 transcriptional regulation was analysed by RNA-seq analyses, which revealed that MAPK12-G4 inhibits oncogenic RAS transformation and trans-activation of NANOG. MAPK12-G4 also reduces CD44 High /CD24 Low population in TNBC cells and downregulates internal stem cell markers, arresting the stemness properties of cancer cells.
Keyphrases
- signaling pathway
- transcription factor
- stem cells
- pi k akt
- oxidative stress
- induced apoptosis
- gene expression
- rna seq
- cell cycle arrest
- dna methylation
- epithelial mesenchymal transition
- crispr cas
- single cell
- ejection fraction
- breast cancer cells
- poor prognosis
- high resolution
- diabetic rats
- long non coding rna
- cell death
- heat stress
- peritoneal dialysis
- drug induced
- water soluble
- embryonic stem cells