IRE1alpha cancer mutants diiferentially regulate Rho family GTPases

IRE1alpha is a multi-domain, ER-transmembrane protein that can mediate the Unfolded Protein Response (UPR). Here, we show that cancer-specific mutations in IRE1alpha can differentially activate the Rho family GTPases to mediate tumor promoting events in mouse keratinocytes. To investigate the roles of cancer-relevant mutations in IRE1alpha during skin carcinogenesis, we generated multicellular spheroids (MCS) of immortalized mouse keratinocytes line C57-T inducibly expressing wildtype (WT) or cancer-relevant IRE1alpha mutations for 24h using doxycycline. We pooled 30 spheroids per cell line per replicate, extracted total cellular RNA, which was used for differential gene expression profiling by RNA-Seq to identify genes potentially regulated by IRE1alpha. Biological triplicates were generated and processed for all conditions. Samples were compared against the parent C57-T cells.