Non-histone chromatin protein in regenerating-rat liver.
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Following the intraperitoneal administration of [3H] tryptophan and [35S] methionine to partially hepatectomized rats, nuclei and chromatin were prepared from the liver. Three protein fractions were prepared from the chromatin by differential salt extractions. The 0.35M NaCl-soluble (0.35 MS) fraction was rich in non-histone chromatin proteins (NHCPs) and the core histones, whereas the 0.35M NaCl-precipitate (2 MS) fraction was composed of the five classes of major histones and two low molecular-weight NHCPs. The 2M NaCl-precipitate (2 MP) fraction was mainly composed of high molecular-weight NHCPs. The incorporation levels were higher in the 0.35 MS and 2 MP fractions than in the 2 MS fraction. In particular, the 2 MP fraction revealed the highest incorporations 14 h after partial hepatectomy (prereplicative phase). The ratio of 3H/35S was also always higher in the 0.35 MS and 2 MP fractions than in the 2 MS fraction. Two chromatin subtractions, 2mM Mg2+-soluble (S 2) and insoluble (P 2) fractions, were prepared after the digestion of chromatin with DNase II. The S 2 fraction was mainly composed of the core histones and high molecular-weight NHCPs, whereas the P 2 fraction was enriched in the five classes of major histones and low molecular-weight NHCPs. The incorporation levels of the above amino acids were always higher in S 2 than P 2. The ratio of 3H/35S was also higher in S 2 than P 2 14 h after partial hepatectomy. In addition, the incorporations into 0.35 MS, 2 MP, and S 2 fractions were considerable even at 0 time. From these findings, it is evident that the high molecular-weight NHCPs were synthesized preferentially in the prereplicative phase, and showed a rapid turnover in normal liver. On the other hand, the incorporation of [14C]leucine into the histone fraction was enhanced during the DNA replication. However, the incorporation levels of the 2 MS fraction were elevated at 14 h and further elevated at 20 h after partialhepatectomy although the levels were lower than in the other two-protein fractions. These results suggest histone synthesis before the onset of DNA replication. In this connection, the quantitative changes in low molecularweight NHCPs, HMG and A 24 proteins were examined by electrophoretic analysis. One of HMG proteins adjacent to H1 histone and A 24 protein increased in the regenerating livers whereas two other HMG proteins, probably HMG 14 and 17, decreased drastically from the prereplicative to the replicative phase.
- 社団法人 日本生化学会の論文
社団法人 日本生化学会 | 論文
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