Multivalent binding of PWWP2A to H2A.Z regulates mitosis and neural crest differentiation

A

Heatmap of significant outliers from pull‐downs analyzed by label‐free MS‐based proteomics with mononucleosomes derived from Skmel‐147 (two independent experiments: SK1 and SK2; see also Vardabasso et al, 2015 for first replicate) or HeLaK (HK) cells stably expressing GFP‐H2A (H2A), GFP‐H2A.Z.1 (Z.1), or GFP‐H2A.Z.2 (Z.2), normalized to H2A. Scale bar: log2‐fold. See also Fig EV1A–E for details on experimental procedure and verification of results, and Datasets EV1, EV2 and EV3 for detailed lists of H2A.Z‐binders.

B

Immunoblotting of SRCAP complex‐specific member ZNHIT1 upon SK‐mel147 GFP, GFP‐H2A, GFP‐H2A.Z.1, or GFP‐H2A.Z.2 mono‐IPs. GFP served as control.

C, D

Immunoblotting of different histone PTMs (C) or PWWP2A (D) upon GFP, GFP‐H2A, GFP‐H2A.Z.1, or GFP‐H2A.Z.2 mono‐IPs. Notice the different sizes of endogenous PWWP2A protein (see also Figs EV1H and EV4C), possibly due to different modifications.

E

Immunoblots of GFP‐H2A.Z.1 or GFP‐PWWP2A mono‐IPs detecting endogenous H3, H2A, or H2A.Z. Notice enrichment of H2A.Z in comparison with H2A in GFP‐PWWP2A pull‐down. See also Fig EV1F–H for generation and characterization of GFP‐PWWP2A cell lines and endogenous pull‐down.

Figure 1. PWWP2A is a novel H2A.Z mononucleosome binder.