Histone Biosynthesis and the Cell Cycle

Hnilica, Lubomir S.; McClure, M. E.; Spelsberg, Thomas C.

doi:10.1007/978-1-4684-1815-6_5

Cited by 9 publications

(4 citation statements)

References 238 publications

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“…Histone biosynthesis takes place at small cytoplasmic polysomes [5] during the DNA synthesis phase [6]. Since histone methylation in vivo is performed by a nuclear enzyme [7], the occurrence of radioactive methyllysines immediately after injection of [3H]lysine implies that the transport of newly synthesized histones into the nucleus must be a rather fast process.…”

Section: Histone Methylation and Cell Cyclementioning

confidence: 99%

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Kinetics of Histone Methylation in vivo and Its Relation to the Cell Cycle in Ehrlich Ascites Tumor Cells

Thomas¹,

Lange²,

Hempel³

1975

Eur J Biochem

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The appearance of methylated lysines in newly synthesized histones from Ehrlich ascites tumor cells was measured during one generation time. Newly synthesized histones were pulse-labeled in vivo by ~- [~H]lysine, and the time course of the uptake of label into monomethyl, dimethyl and trimethyllysine from gel-electrophoretically isolated histones F2al (H4) and F3 (H3) was followed.Methylation starts immediately after histone biosynthesis. It proceeds, however, more slowly than histone synthesis. Both the rate of methylation and the mechanism of methylation in F3 and F2al histones differ. F3 methylation can be described by a first-order reaction, i.e. the reaction rate depends only on the concentration of free methylation sites available. Rate constants of approximately 0.21 h-' were found for all three methylation steps.Methylation in the F2al histone proceeds more slowly than in F3. The dimethylation step in this fraction can be described by a zero-order reaction with a rate constant which is the reciprocal of the duration of the DNA synthesis phase. Alternatively this step could be correlated with the transition of the cells from the S phase into the G2 phase.By the end of one generation time all methylation sites in all F2al and F3 molecules are occupied by methyl groups at a ratio of about 1 : 3 : 1 for monomethyl, dimethyl and trimethyllysine in the F3 histone. In the F2al molecule the methyllysines consist mainly of dimethyllysine.The lysine in position 20 in the arginine-rich histone fraction F2al (histone H4) and the lysines in positions 9 and 27 in the F3 histone (histone H3) can be methylated in vivo [l]. The methylation may lead to NE-monomethyllysine [2], ,V-dimethyllysine [3], and XE-trirnethyllysine [4]. In contrast to histone synthesis, which takes place at small cytoplasmic polysomes [5] during the DNA synthesis (S) phase [6], histone methylation is performed in the nucleus [7].There is no general agreement in the literature about when methylation takes place during the mitotic cycle. After partial hepatectomy histone methylation has been reported during [8] and for a significant time after [9] DNA synthesis. With synchronized HeLa cells histone methylation was found throughout Part of this work was presented at the 9th FEBS Meeting. Budapest, 1974. Abbreviations. Lys(Me), methyllysines; Lys(Mel), N' -monomethyllysine ; Lys(Me,), NE -dimethyllysine; Lys(Me,), NE-trimethyllysine. Histones F2a 1 and F3 correspond to histones H4 and H3, respectively, in the nomenclature agreed at a recent CIBA conference (in press).-~ the cell cycle with a maximum ranging from late S phase through G2 to mitosis [lo]. Experiments with Chinese hamster ovary cells suggested methylation of the arginine-rich histones during late S and G2 phases [ll].In a previous paper we have shown that there is not a rapid turnover of the histone-bound methyl groups [12] in contrast to the relatively rapid metabolism of some of the histone-bound acetate and phosphate groups [I 31. The present communication deals with the kinetics of ...

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Section: Histone Methylation and Cell Cyclementioning

confidence: 99%

“…The methylation may lead to NE-monomethyllysine [2], ,V-dimethyllysine [3], and XE-trirnethyllysine [4]. In contrast to histone synthesis, which takes place at small cytoplasmic polysomes [5] during the DNA synthesis (S) phase [6], histone methylation is performed in the nucleus [7].…”

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Kinetics of Histone Methylation in vivo and Its Relation to the Cell Cycle in Ehrlich Ascites Tumor Cells

Thomas¹,

Lange²,

Hempel³

1975

Eur J Biochem

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“…Elucidating the manner in which the expression of viral information, integrated within the host cell genome, is regulated is central to understanding the mechanism by which maintenance of the transformed state is acieved. Since chromosomal protein --historic and non-historic chromosomal proteins --play a key role in the regulation of transcription (Allfrey et al, 1963;Huang et al, 1965;Gilmour et al, 1970;Kleinsmith et al, 1970;Spelsberg et al, 1970;Teng et al, 1971;Kostraba et al, 1973;Paul et al, 1973;Stein et al, 1972aStein et al, , b, 1974bStein et al, , 1975 as well as in the determination of chromatin structure (Allfrey, 1971;Clark et al, 1971;Hnilica et al, 1971;Hewisch et al, 1973;Noll, 1974;Kornberg, 1974;Kornberg et al, 1974;Rill et al, 1973;Krause et al, 1974c;Olins et al, 1974;Stein et al, 1974;Weintraub et al, 1974a), it is reasonable to anticipate the potential involvement of these proteins in mediating viral-induced modifications in gene readout.…”

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The effect of SV40 transformation on the chromosomal proteins of 3T3 mouse embryo fibroblasts

Krause

Noonan

Kleinsmith

et al. 1976

Cell Differentiation

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The composition and metabolism of chromosomal proteins-histones and nonhistones chromosomal proteins-were examined in normal and SV40 transformed 3T3 mouse cells. Variations were observed, many of which were similar to those previously reported for normal and SV40 transformed W138 human diploid fibroblasts. The possible implications of these viral induced changes in the protein component of the genome for the phenotypic modifications which occur in transformed cells are discussed.

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“…Several lines of evidence suggest that the proteins associated with the genome play such a regulatory role in eukaryotic cells (17,18). Histones have been shown to be involved in the repression of I;DNAdependent RNA synthesis (19,20), maintenance of chromosome structure (21), and cytodifferentiation (22). Nonhistone chromosomal proteins may recognize defined gene loci and thus mediate specificity of gene read-out (18,(23)(24)(25)(26)(27)(28)(29).…”

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Gene expression in mouse neuroblastoma cells: properties of the genome.

Zornetzer¹,

Stein²

1975

Proc. Natl. Acad. Sci. U.S.A.

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Chromatin was prepared from isolated nuclei of proliferating and differentiated cultures of C1300 mouse neuroblastoma cells. Differentiation (2-8), biochemical (4-7, 9-14), and electrical properties characteristic of neurons (15,16).The complex changes that accompany transition from the proliferating to the differentiated state may reflect alterations in the expression of information contained within the genome. One might anticipate modifications, therefore, in the macromolecules that comprise the genome and interact with DNA to regulate its function. Several lines of evidence suggest that the proteins associated with the genome play such a regulatory role in eukaryotic cells (17,18). Histones have been shown to be involved in the repression of I;DNAdependent RNA synthesis (19,20), maintenance of chromosome structure (21), and cytodifferentiation (22). Nonhistone chromosomal proteins may recognize defined gene loci and thus mediate specificity of gene read-out (18,(23)(24)(25)(26)(27)(28)(29). The nonhistone proteins have been shown to direct the tissue specific transcription of globin genes (41-43) and the cell cycle stage-specific transcription of histone genes (44,45). In the present study we have compared the histones and nonhistone chromosomal proteins associated with the genome in proliferating and differentiated mouse neuroblastoma C1300 cells. The possibility of a functional relationship between chromosomal proteins and differences observed in the transcriptional activity of chromatin from proliferating and differentiated mouse neuroblastoma cells is discussed. MATERIALS AND METHODSThe neuro 2a line of mouse neuroblastoma C1300 cells was grown in monolayer culture in 1-liter glass Blake flasks in Eagle's minimum essential medium (MEM) supplemented with 10% fetal calf serum in a moist CO2 incubator. To induce differentiation, cells were plated in MEM with 20% fetal calf serum and incubated for 24 hr. The monolayers were then washed with MEM and subsequently maintained for 24 hr in either serum-free MEM or in MEM with 20% fetal calf serum and 1 mM dibutyryl cyclic AMP. Proliferating cultures were treated in a similar manner, but 20% fetal calf serum was present during both 24-hr incubation periods.Nuclei and chromatin were prepared from mechanically harvested cells at 40. Details of the procedures have been described (30,31). Isolated nuclei were free of cytoplasmic contamination when examined by phase contrast microscopy. Little difference was observed in the patterns of proteins fractionated electrophoretically from either nuclei or isolated chromatin when comparisons were made with and without the inclusion of 50 Mg/ml of L-1-tosylamide-2-phenyl ethyl chloromethyl-ketone (TPCK) in the isolation and washing media to inhibit proteolysis (32). The histone:nonhistone chromosomal protein ratios in chromatin from proliferating and differentiated cultures were 0.73 and 0.79, respectively.The ability of chromatin from proliferating and differentiated cells to support RNA synthesis in vitro was determ...

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Histone Biosynthesis and the Cell Cycle

Cited by 9 publications

References 238 publications

Kinetics of Histone Methylation in vivo and Its Relation to the Cell Cycle in Ehrlich Ascites Tumor Cells

Kinetics of Histone Methylation in vivo and Its Relation to the Cell Cycle in Ehrlich Ascites Tumor Cells

The effect of SV40 transformation on the chromosomal proteins of 3T3 mouse embryo fibroblasts

Gene expression in mouse neuroblastoma cells: properties of the genome.

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