Project description:O-linked β-N-acetylglucosamine (O-GlcNAc), a critical post-translational modification (PTM) predominantly found in the nucleus, plays a substantial role in regulating gene expression by modulating transcription factors (TFs) activity. Herein, by utilizing a pulldown screening approach termed as tandem array of transcription factors response elements (catTFREs), we unravel the profound impact of fluctuating levels of O-GlcNAcylation on the DNA binding efficiency of endogenous TFs. The proteins were enriched for label-free quantitative proteomics by LC-MS/MS.
Project description:To find new transcription and splicing factors associated with the metastatic phenotype we used a concatenated tandem array of consensus transcription factors response elements (catTFRE) on nuclear extracts of colon cancer cell lines. We used two different isogenic pairs of colorectal metastatic cells, KM12SM/KM12C and SW620/480. Proteins were pulled-down using biotinylated DNA sequences and streptavidin beads and analyzed by LC-MSMS.
