Fig. 4: GAND–associated missense variants L180P, R414Q, C420S, and C420R disrupt binding to nucleosome remodeling and deacetylase (NuRD) components CHD3/4/5 or MBD2/3. | Genetics in Medicine

Fig. 4: GAND–associated missense variants L180P, R414Q, C420S, and C420R disrupt binding to nucleosome remodeling and deacetylase (NuRD) components CHD3/4/5 or MBD2/3.

From: GATAD2B-associated neurodevelopmental disorder (GAND): clinical and molecular insights into a NuRD-related disorder

Fig. 4

a FLAG-tagged MBD2 or MBD3 were coexpressed with HA-GATAD2B (wild-type [WT] or L180P) by in vitro transcription–translation (IVT) in a rabbit reticulocyte lysate. b The FLAG-tagged C1–C2 domains of CHD3, CHD4, or CHD5 were coexpressed with HA-GATAD2B (WT, R414Q, C420R, C420S, and G406R) in IVTs. In all experiments, FLAG-fusion proteins were immobilized on αFLAG affinity beads and used as baits to pull down the coexpressed HA-GATAD2B. As a negative control, wild-type (WT) HA-GATAD2B was added to beads to which no FLAG-fusion protein had been immobilized. In each case, 10% of inputs and 50% of elutions were loaded on an sodium dodecyl sulfate–polyacrylamide gel electrophoresis (SDS-PAGE) and proteins were detected by western blot, using αHA and/or αFLAG antibodies. # degradation product of FLAG-MBD2.

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