Histone h4 is a highly conserved structural component of the nucleosome subunit of chromatin. The activation of chromatin is accompanied by changes in structure which may be caused by histone modification or by interactions of specific non-histone proteins, or both. Histone H4 can be modified by acetylation and this modification has been correlated with chromosome assembly and with transcription. We have now tested these correlations by studying H4 acetate content as a function of the cell cycle using the naturally synchronous cell cycle in Physarum polycephalum. The results show two clear correlations: (1) tetra-acetylated H4 correlates with transcription; (2) highly acetylated H4 (2 to 4 acetates per molecule) is inversely correlated with H1 phosphorylation and initiation of chromosome condensation in prophase. The results are consistent with turnover of di-acetylated H4 during chromosome assembly in S phase.