Yeast contains multiple forms of histone acetyltransferase.
We have assayed several methods to quantitatively recover yeast histone acetyltransferases in an attempt to study the multiplicity of enzymatic activities. Two methods, namely (NH4)2SO4 precipitation and salt dissociation of chromatin in 0.5 M NaCl, yielded convenient preparations of total histone acetyltransferases. DEAE-Sepharose chromatography of the crude extracts resulted in the separation of three peaks of activity when total yeast histones were used as substrate. However, the scanning of the enzymatic activity toward individual histones along the chromatography, achieved by determining the specific activity of the individual histones after incubating whole histones and [14C]acetyl-CoA with the chromatographic fractions, yielded four peaks. The first two peaks showed specificity toward H2B and H3, respectively. Although they partially overlapped, rechromatography on cation exchangers allowed us to resolve the two activities, and several criteria were used to prove that they correspond to different enzyme molecules. The last two peaks were H4-specific, but the present data suggest that one of the activities is chromatin-bound, whereas the other surely corresponds to the cytoplasmic B-form of the enzyme. The enzyme specific for yeast H2B acetylates chicken erythrocyte H2A, rather than H2B. The detected multiplicity of yeast histone acetyltransferases may correspond to the multiplicity of roles proposed for histone acetylation.
López-Rodas
G
G
Department of Biochemistry and Molecular Biology, Faculties of Sciences, University of Valencia, Spain.
Tordera
V
V
Sánchez del Pino
M M
MM
Franco
L
L
D000107:Acetylation; D000123:Acetyltransferases / Q000378:metabolism*; D002843:Chromatin / Q000201:enzymology; D002852:Chromatography, Ion Exchange; D004795:Enzyme Stability; D051548:Histone Acetyltransferases; D006655:Histone Deacetylases / Q000302:isolation & purification / Q000378:metabolism; D006657:Histones / Q000302:isolation & purification; D007527:Isoenzymes / Q000302:isolation & purification / Q000378:metabolism*; D007700:Kinetics; D012441:Saccharomyces cerevisiae / Q000201:enzymology*; D029701:Saccharomyces cerevisiae Proteins; D013379:Substrate Specificity
D002843:Chromatin; D006657:Histones; D007527:Isoenzymes; D029701:Saccharomyces cerevisiae Proteins; D000123:Acetyltransferases; D051548:Histone Acetyltransferases; D006655:Histone Deacetylases