Fig. 7

Tyrosine residue 572 of ZMYND11 is functionally important and essential for specifically reading SDMA on HNRNPA1. a Schematic representation of the MYND domain of ZMYND11. Shown is MYND domain organization along with potential candidate amino acid residues likely to be involved in binding with HNRNPA1. MYND domain conserved across species is shown. b Two views of the crystal structure (PDB:5HDA) for the MYND domain of ZMYND11 complexes with EBNA2 (blue). Potential candidate amino acid residues responsible for recognition with HNRNPA1 are show in sticks. c Co-immunoprecipitation assay in HEK293T cells expressing T7-tagged HNRNPA1 and V5-tagged ZMYND11 or the indicated mutants. d Pull-down assays demonstrated protein-protein interactions between HIS-tagged MYND domain of ZMYND11 or the mutants and the biotinylated peptide containing R194 SDMA. Representation and quantification of colony-forming (e), cell proliferation (f), migration (g), and invasion (h) of 22Rv1 cells overexpressing ZMYND11 or the Y572A mutant. Scale bar, 400 μm. *P < 0.05, **P < 0.01, ***P < 0.001, were examined by two-tailed Student’s t test. PKM splicing assay (i) and immunoblots for the indicated PKM isoforms (j) were performed in 22Rv1 cells ectopically expressing ZMYND11 or the Y572A mutant. k Relative glucose uptake and lactate production were measured in 22Rv1 cells with an ectopical expression of ZMYND11 or Y572A mutant. ns, not significant, *P < 0.05, **P < 0.01, ***P < 0.001, two-tailed Student’s t test. l Fluorescence colocalization microscopy analysis of ZMYND11 or the Y572A mutant with HNRNPA1 in 22Rv1 cells treated with sodium arsenite or control. Note that stress-induced cytoplasmic accumulation of HNRNPA1 was substantially attenuated by wild-type ZMYND11, but not the Y572A mutant. Arrows, stress granules in cells. Mean ± SD, was assessed using two-tailed Student’s t test. *** represents p < 0.005. Scale bars, 25 μm