ATAC-seq profiling of mutated MORC2 overexpression in MORC2-knocked-out HEK293T cells

We report an in vitro reconstitution of full-length MORC2, the most commonly mutated MORC member, linked to various cancers and neurological disorders. MORC2 possesses multiple DNA binding sites that undergo structural rearrangement upon DNA binding. MORC2 locks onto the DNA using its C-terminal domain (CTD) and acts as a sliding clamp. A conserved phosphate-interacting motif within the CTD was found to regulate ATP hydrolysis and cooperative DNA binding. Importantly, MORC2 mediates chromatin remodelling via ATP hydrolysis-dependent DNA compaction, regulated by the phosphorylation state of its CTD. Overall design: MORC2 knockout cell lines were contructed from the HEK293T cell line using two different sgRNAs. Plasmid were then used to restore MORC2 expression, either of WT MORC2 or of two mutated MORC2 variants. ATAC-seq profiling was conducted in duplicate on knock-out HEK293T cell lines with (i) a plasmid carrying MORC2-GFP (WT), (ii) a plasmid carrying MORC2 phosphodead-GFP (S6A), and (iii) a plasmid carrying MORC2 phosphomimetic-GFP (S6D).