Structure-Function analysis of a novel DNA-binding domain of the histone acetyltransferase MOZ/KAT6ATools Schoenfeld, Jonas (2019) Structure-Function analysis of a novel DNA-binding domain of the histone acetyltransferase MOZ/KAT6A. MRes thesis, University of Nottingham.
AbstractIn this study, we focussed on the poorly characterised NEMM domain comprised within the N-terminal 200 amino acids of the Monocytic Leukemia Zinc Finger Protein (MOZ/KAT6A). MOZ is an important developmental regulator and involved in several diseases such as neurodevelopmental disorders and cancers. The NEMM domain of MOZ was recently found to be essential for induction of acute myeloid leukemia (AML) by the pathological fusion protein MOZ-TIF2. Structural predictions suggest that this region forms a potential tandem winged-helix domain (WHD) as found in a subfamily of DNA-binding proteins. Consistent with this, EMSA experiments indicated that the NEMM formed complexes with random dsDNA fragments in vitro. Mapping analysis showed that the first predicted winged-helix domain (pWHD1) preferentially binds dsDNA sequences containing CpG motifs, whereas the preference for pWHD2 was not defined. However, the data suggest that the tandem WHDs may form a composite DNA interaction-surface. Preliminary mutational analysis defined a lysine-rich stretch as necessary for DNA-recognition by pWHD1. Large-scale purification of MOZ pWHD1 free from contaminating DNA was also optimised, generating high yields of homogenous protein for protein crystallisation trials.
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