Systematic phenotypic characterization and integrated analysis show heterogeneous neomorphic activities of different missense mutant p53 proteins

The phenotypic and transcriptomic data on ten normal breast epithelial cell lines each expressing a distinct missense mutant p53 protein, and their systems-level analysis through ChIP-Seq and RNA-seq defines the molecular basis for phenotypic heterogeneity imparted by the different missense mutant p53 proteins. The focus of the study is to gain mechanistic insight on the neomorphic function of mutant p53 proteins and their manifestation into phenotypic heterogeneity in the context of a TNBC model. Overall design: Examining the transcriptomics of 10 different p53 missense mutant protein expressing MCF10A cell lines using RNA-seq, and DNA binding and promoter occupancy of 5 p53 mutant MCF10A cell lines using ChIP-Seq. The missense mutant p53 proteins have a V5 tag at the C-term and for ChIP, the mutant p53 proteins were pulled down using the V5 antibody. For the ChIP-seq, the MCF10A cells with endogenous wild type p53 served as the negative control and the cells expressing WT p53 with the V5 tag at C-term as the positive control.