Phosphorylation and dephosphorylation of histone V (H5): controlled condensation of avian erythrocyte chromatin. Appendix: Phosphorylation and dephosphorylation of histone H5. II. Circular dichroic studies

Abstract

During avian erythropoiesis, the blast cells of the bone marrow mature into polychromatic erythrocytes (late stages known as reticulocytes) and then into mature red blood cells. When chickens are made anemic, the proportion of immature cells in the anemic bone marrow increases dramatically. The level of the lysine-rich histones, H1 and H5, was constant in the blood and bone marrow of normal and anemic chickens. This implies that H5 replaces H1 quantitatively. Urea-aluminum-lactate starch gel electrophoresis of H5 from these sources shows that the degree of phosphorylation of H5 is proportional to the number of immature cells. About 70% of the H5 from the most immature bone marrow is phosphorylated, while 50% of the H5 from anemic blood is phosphorylated and H5 in normal blood is almost completely devoid of phosphate. When immature cells of the anemic bone marrow are incubated in the presence of 32Pi and [3H]lysine and [3H]arginine, extensive 32P incorporation is found in the phospho species. A minimum of 9 phosphorylated components were demonstrated by starch gel electrophoresis. The incorporation of 3H is time dependent. After 1.5 h of labeling, 3H is found in H5 containing 0, 1, 2 and 3 phosphates. Newly synthesized H5 may become progressively phosphorylated; at the terminal stage of development, the phosphorylated H5 is probably completely dephosphorylated. These events may be important in controlling the timing of chromatin condensation.