The heterogeneity of the non‐histone chromatin proteins from mouse tissues

MacGillivray, A.J.; Carroll, Dana; Paul, John

doi:10.1016/0014-5793(71)80536-7

Cited by 87 publications

(17 citation statements)

References 25 publications

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“…These proteins possess tissue specificity (10)(11)(12)(13)(14)(15)(16)(17) and are capable of interacting with DNA to selectively modify transcription (18,19). In HeLa cells, nonhistone chromosomal proteins are actively synthesized throughout the cell cycle (20), including the mitotic phase (21).…”

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Role of Nonhistone Chomosomal Proteins in the Restriction of Mitotic Chromatin Template Activity

Stein

Farber

1972

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

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RNA synthesis virtually ceases during mitosis in eukaryotic cells. This phenomenon is explainable, at least in part, by the reduced template activity for RNA synthesis of mitotic chromatin. The restriction in template activity can be accounted for by the presence in the chromatin of mitotic-specific nonhistone proteins. Chromatin reconstituted by gradient dialysis from the pooled histones of S-phase and mitotic chromatin and the nonhistone proteins of mitotic chromatin had a template activity similar to that of native mitotic chromatin, and lower than native S-phase chromatin or chromatin reconstituted with the pooled histones and the S-phase and nonhistone proteins. The template activity was identical for chromatin reconstituted from the pooled nonhistone proteins and either the histones from S-phase or mitotic chromatin. These results are supported with data showing that the procedure of reconstitution produces a chromatin indistinguishable in its protein composition both qualitatively and quantitatively from its native counterpart.Macromolecular metabolism is strikingly altered in eukaryotic cells during mitosis. RNA synthesis virtually ceases (1-4), and the rate of protein synthesis is significantly reduced (3, 5). Johnson and Holland (6) reported that mitotic chromatin has a reduced template activity for RNA synthesis. We recently confirmed this observation and have shown that this restriction of chromatin template activity may be produced by the chromosomal protein fraction specific for mitotic chromatin (7). The template activity of chromatin reconstituted by gradient dialysis from purified HeLa cell DNA and the chromosomal proteins isolated from Colcemidarrested metaphase cells was considerably less than that of chromatin reconstituted from S-phase chromosomal proteins. The degree of template restriction was proportional to the amount of mitotic chromosomal proteins in the reconstituted chromatin. This finding led us to suggest that the inhibition of template activity of mitotic chromatin might be accounted for by a constituent of the chromosomal protein fraction that binds to DNA during mitosis and thereby prevents transcription by RNA polymerase (7).Recent studies from several laboratories have indicated that the nonhistone chromosomal proteins may play a significant role in the control of gene expression in eukaryotic cells (8,9). These proteins possess tissue specificity (10-17) and are capable of interacting with DNA to selectively modify transcription (18,19 Fla. 32601 in the nonhistone proteins have been described in various phases of the HeLa cell cycle (20,22).In this report we show that nonhistone chromosomal proteins may be responsible for the inhibition of template activity of mitotic chromatin. MATERIALS AND METHODSExponentially growing HeLa S3 cells were maintained in suspension culture in Joklik-modified Eagle's Minimal Essential Medium supplemented with 3.5% (each) of calf and fetal-calf serum. Synchronization of the cells was produced (20) by 2 cycles of 2 mM thymidine block...

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confidence: 99%

Role of Nonhistone Chomosomal Proteins in the Restriction of Mitotic Chromatin Template Activity

Stein

Farber

1972

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

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“…Analyses of nuclear acidic proteins in various human normal and leukaemic cell lines gain importance since it has been shown that some of these proteins are specific in different tissues and in different species [15,25,38,41,42]. It has been derived from transcriptional studies that nonhistone proteins are probably involved in positive control mechanisms for gene expression [2,7,13,18,22,31,35].…”

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Comparative studies on phenol-soluble nonhistone chromatin proteins in normal and leukaemic human leukocytes

et al. 1979

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“…Thus fraction Hap 1 consists of histones whereas Hap 2-4 are fractions of chromatin non-histone proteins. The Hap 1 fraction gave a quantitative recovery of all histone fractions as is obtained by acid-extraction techniques ; there was no loss of histone H1 [14]. Polynucleotide material was removed from each of the protein fractions by centrifugation in CsCl/urea gradients as described elsewhere.…”

Section: Characterisation Ofthe Labelled Productsmentioning

confidence: 99%

“…["PIHAP 2 fraction was prepared and subjected to density gradient centrifugation as described in Fig.2. Fractions corresponding to (A) the top (1 -6), (B) middle (7)(8)(9)(10)(11)(12) and (C) bottom (13)(14)(15)(16)(17)(18)(19)(20) and (B) bottom ( 1 -1) were calculated to give an estimate of the radioactivity incorporated into proteinbound and free poly(ADP-Rib) respectively. The arrows indicate the time at which nicotinamide (50 mM) was added and the subsequent decrease in acid-insoluble radioactivity at the top (0---) and bottom (&a) of the gradients was determined associated with both protein-bound and free poly-(ADP-Rib).…”

Section: Synthesis Of Free Poly(adp-rib)mentioning

confidence: 99%

The Modification of Nuclear Proteins by ADP‐ribosylation

Rickwood

MacGillivray

Whish

1977

European Journal of Biochemistry

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When incubated in vitro purified mouse nuclei incorporate NAD into poly(ADP-Rib). Analysis of the product on CsCl/urea gradients showed that a large proportion of the poly(ADP-Rib) was not attached to protein. The free poly(ADP-Rib) did not appear to arise through degradation and its chain length was significantly longer than the poly(ADP-Rib) attached to proteins. Fractionation of the proteins on hydroxyapatite revealed that tissue-specific modification of both the histones and non-histone proteins, had occurred. In the case of one protein species there was evidence for the existence of several forms with different numbers of ADP-Rib residues.It is now more than ten years since the synthesis of poly(ADP-Rib) in isolated nuclei was first demonstrated; it was shown that when nuclei were incubated in vitro with labelled NAD the radioactivity was incorporated exclusively into poly(ADP-Rib) [l]. More recently evidence has been presented to show that this polymer is also synthesized in vivo [2]. Poly(ADP-Rib) is a polynucleotide-like molecule but, in contrast to the other polynucleotides of the cell, its functional role in the cell remains unclear. It has been reported that poly(ADP-Rib) is attached to the nuclear proteins (for a review see [3]) and as such it could modify the characteristics of the proteins to which it is bound, in the same way as phosphorylation and acetylation can also modify the interaction of histones with DNA. Two problems emerge from the evidence presently available. The first concerns which proteins of the nucleus act as acceptors for the poly(ADP-Rib) molecules. So far only the histones have been identified with certainty as being modified by the polymer [4-61, but with the improved techniques now available (see [7]) it is possible to examine the role of most of the nuclear proteins in this process. The second problem relates to the large number of conflicting reports as to the rate of synthesis of poly(ADP-Rib) at different times of the cell cycle [8] and the difficulty of correlating polymer production and its modification of nuclear proteins with particular events in the cell cycle or with changes in gene activity. For some years these laboratories have been involved in developing procedures for the preparation and analysis of nuclear proteins (see [7,9,10]) and in this paper we describe the application of some of our techniques to study the modification of nuclear proteins by poly-(ADP-Rib) in a number of mouse cells.

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The heterogeneity of the non‐histone chromatin proteins from mouse tissues

Cited by 87 publications

References 25 publications

Role of Nonhistone Chomosomal Proteins in the Restriction of Mitotic Chromatin Template Activity

Role of Nonhistone Chomosomal Proteins in the Restriction of Mitotic Chromatin Template Activity

Comparative studies on phenol-soluble nonhistone chromatin proteins in normal and leukaemic human leukocytes

The Modification of Nuclear Proteins by ADP‐ribosylation

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