Fig 6. Amino acid residues 31 and 32 are essential for the function of H3V^CTG.

(A) Schematics represent various point-mutant strains constructed by changing amino acid residues at positions 31, 32, and/or 80 of variant histone H3 to that of the canonical histone H3. (B) SC5314, H3V^CTG null mutant LR108, H3V^CTG complemented strain LR109, canonical histone H3 mutant LR142 (hht21/hht21), point-mutant strains at position 31 (RS103), 32 (RS105), 80 (RS107), and point-mutant strain at positions 31 and 32 (RS109) were allowed to form biofilms in YPDU for 48 hours at 37 °C; wells were washed to remove the nonadherent cells and photographed. (C) Biomass dry weights of the wild-type, H3V^CTG null mutant, H3V^CTG complemented, and hht21/hht21 strains grown in YPDU at 37 °C. The data underlying this figure can be found in S2 Data. (D) Biomass dry weights of the wild-type, H3V^CTG null mutant, H3V^CTG complemented, and point-mutant strains grown in YPDU at 37 °C. The data underlying this figure can be found in S2 Data. (E) SC5314, H3V^CTG null mutant LR108, H3V^CTG complemented strain LR109, canonical histone H3 mutant LR142 (hht21/hht21), point-mutant strain at position 31 (RS103), 32 (RS105), 80 (RS107), and point-mutant strain at position 31 and 32 (RS109) were spotted on synthetic dextrose medium complemented with essential amino acids (CM) agar plates, and CM+NAG (synthetic dextrose with 1 mM N-acetyl glucosamine) agar plates, and incubated for 3 days at 37 °C. (F) The extent of filamentation was monitored for the indicated strains by growing colonies from single cells on CM medium at 37 °C. (G) SC5314, H3V^CTG H3 null mutant LR107, H3V^CTG complemented strain LR109, point-mutant strains at position 31 (RS103), 32 (RS105), 80 (RS107), point-mutant strain at positions 31 and 32 (RS109), and canonical histone H3 mutant LR142 (hht21/hht21) were allowed to form filaments in liquid YPD with 10% FBS at 37 °C. The proportion of hyphal cells formed by these strains are plotted. The data underlying this figure can be found in S2 Data. CM, complete media; CM+NAG, complete media supplemented with N-acetyl glucosamine; FBS, fetal bovine serum; ns, not significant; YPDU, yeast peptone dextrose supplemented with uridine.