Biochem. J. (2014) 461 (323–334) (Printed in Great Britain) doi:10.1042/BJ20140374
PRDM proteins have emerged as important regulators of disease and developmental processes. To gain insight into the mechanistic actions of the PRDM family, we have performed comprehensive characterisation of a prototype member protein, the histone methyltransferase PRDM9, utilising biochemical, biophysical and chemical biology techniques. We report the first known molecular characterisation of PRDM9-methylated recombinant histone octamer and the identification of new histone substrates for the enzyme. A single Cys321Pro mutant of the PR/SET domain was demonstrated to significantly weaken PRDM9 activity. Additionally, we have optimised a robust biochemical assay amenable to high throughput screening to facilitate the generation of small molecule chemical probes for this protein family. This work has provided valuable insight into the enzymology of an intrinsically active PRDM protein.
Fast-Track Joint-Council Office Development Programme Grant [grant number 1134c001] from A*STAR.