Genome-wide changes in active RNAPII occupancy in the presence of mutated TFIIS (TFIISmut)

The expresion of dominant negative version of Arabidopsis TFIIS (TFIISmut) in plants lacking funtional TFIIS leads to severe growth defects. The genome-wide analysis of changes in active RNAPII occupancy upon TFIISmut expression was determined using ChIP-seq in order to better understand the biological importance of TFIIS. 10DAS seedlings of transgenic lines carring inducible GFP-TFIIS or GFP-TFIISmut transgenes were studied by ChIP-seq following plants exposure to 24h b-estradiol or EtOH (mock) induction. Overall design: Chromatin DNA immunoprecipitation was performed to determine active RNAPII occupancy using antbodies directed agains S2P-RNAPII or S5P-RNAPII. RNAPII occupancy was studied in 10-day old transgenic plants with inducible expression of GFP-TFIIS or GFP-TFIISmut fusion proteins and analysed comparatively with uninduced plants.