Alterations in amounts and covalent modifications of low-molecular-weight chromosomal proteins in Chinese hamster ovary cells during polyamine depletion.
The effect of polyamine depletion on phosphorylation and ADP-ribosylation of low-Mr chromosomal proteins was studied in intact, mutant Chinese hamster ovary cells (CHO-P22) devoid of ornithine decarboxylase activity. When starved of polyamines for 6 days, severe polyamine deficiency develops and the cells gradually stop growing. The rate of DNA synthesis was retarded to 16% of the control value and to 29% in density-inhibited cells. The synthesis of high-mobility-group (HMG) proteins was decreased by 65% in polyamine-depleted cells and by 40% in density-inhibited cells. The synthesis of core histones was decreased by 40% both in polyamine-depleted and density-inhibited cells. In polyamine-depleted cells the molar ratio of the higher-Mr HMG proteins (HMG 1 + 2) to the lower-Mr HMG proteins (HMG 14 + P) was about one-half of that found in cells grown in the presence of putrescine or in density-inhibited cells. In contrast to HMG proteins, no major differences were found in the content of core histones in these cell populations. In the perchloric acid-soluble fraction of nuclear proteins, 32P was incorporated mainly into histone H1, HMG P and a protein migrating more slowly than HMG 1 (protein P1). Specific changes in the 32P-labeling and migration of a number of protein bands, including histone H1, was observed in polyamine-depleted cells as compared to cells grown in the presence of putrescine or to density-inhibited cells. ADP-ribosylation experiments using [3H]adenosine showed a different pattern of label distribution; the higher-Mr HMG proteins from polyamine-depleted cells contained about one-half the amount of label found in the proteins from control cells. The lower-Mr HMG proteins and histone H1 were the preferentially labeled proteins in polyamine-depleted cells. Labeling of core histones with [32P]orthophosphate or [3H]adenosine did not differ markedly in the two cell populations. The results obtained using intact polyamine auxotrophic cells indicated that polyamine depletion is connected with more severe alterations in amounts and covalent modifications (phosphorylation and ADP-ribosylation) of HMG chromosomal proteins and histone H1 than core histones.
D000246:Adenosine Diphosphate Ribose / Q000378:metabolism; D000818:Animals; D002460:Cell Line; D002467:Cell Nucleus / Q000378:metabolism; D006224:Cricetinae; D003412:Cricetulus; D004261:DNA Replication; D005260:Female; D006609:High Mobility Group Proteins / Q000096:biosynthesis* / Q000235:genetics; D006657:Histones / Q000096:biosynthesis* / Q000235:genetics; D008970:Molecular Weight; D009955:Ornithine Decarboxylase / Q000172:deficiency; D010053:Ovary; D010766:Phosphorylation; D011073:Polyamines / Q000378:metabolism*
D006609:High Mobility Group Proteins; D006657:Histones; D011073:Polyamines; D000246:Adenosine Diphosphate Ribose; D009955:Ornithine Decarboxylase