Transcriptomics study of a temperature sensitive mutant (E170K, shot2) in CstF77, a subunit of 3' end mRNA maturation complex in Arabidopsis thaliana

To obtain transcriptome-wide effects of shot2 mutation, we performed RNA-Seq analysis of comparing hot1-4 shot2 double mutant with hot1-4 (control with a dominant-negative mutation in HSP101) in the absence and presence of a heat treatment (38 °C/3h). Heat treatment causes transcriptional readthrough events in hot1-4 shot2, specifically in highly heat-induced genes, including the toxic hot1-4 gene. In addition, failure of correct transcript processing leads to reduced accumulation of many HSP RNAs and proteins, suppressing heat sensitivity of the hot1-4 mutant, due to reduction of the toxic mutant HSP101 protein. Overall design: hot1-4 and hot1-4 shot2 seedlings grown on agar plates until stage 1.02 were treated with or without heat stress at 38 °C for 3h followed by 22 °C for 1h. Three biological replicates each of four sample groups (12 total samples)