Tissue-specific chromosomal non-histone protein interactions with DNA.

Wang, S; Chiu, Jen-Fu; Klyszejko-Stefanowicz, L; Fujitani, Hideo; Hnilica, Lubomir S.

doi:10.1016/s0021-9258(17)33763-8

Cited by 42 publications

(5 citation statements)

References 24 publications

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“…These tightly bound non-histone chromosomal proteins have been shown to be (1) bound to active genes (Gates & Bekhor, 1980; Norman & Bekhor, 1981), (2) bound to middle repetitive DNA sequences (Razin et al, 1979), (3) located in the active diffuse chromatin (Wang et al, 1976), (4) in the nuclear matrix (Berezney & Coffey, 1977;Barrack & Coffey, 1982), and (5) capable of enhancing the binding of RNA polymerase enzyme to DNA (Bekhor & Samal, 1977). These proteins have also been reported to serve as acceptor sites for other steroid receptors (Perry & Lopez, 1978;Hamana & Iwai, 1978;Klyzsejko-Stefanowicz et a!., 1976;Ruh et al, 1981;Ruh & Spelsberg, 1983).…”

Section: Discussionmentioning

confidence: 99%

“…Thus, proteins in the CP-3 fraction appear to contain "acceptor activity" which endow DNA with the capacity for nativelike binding of PR. Similar fractions of tightly bound, non-histone proteins are associated with the nuclear matrix (Berezney & Coffey, 1977; Barrack & Coffey, 1980), with active-diffuse chromatin fractions (Wang et al, 1976) and with active structural genes (Bekhor & Mirell, 1979;Gates & Bekhor, 1980; Norman & Bekhor, 1981; Mirell & Bekhor, 1982). This same class of proteins has been reported to enhance transcription of DNA by increasing the binding of RNA polymerase to DNA (Bekhor & Samal, 1977).…”

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confidence: 84%

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Reconstitution of nativelike nuclear acceptor sites of the avian oviduct progesterone receptor: evidence for involvement of specific chromatin proteins and specific DNA sequences

Spelsberg¹,

Gosse

Littlefield

et al. 1984

Biochemistry

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A specific fraction of avian oviduct chromosomal proteins can be reannealed to pure avian DNA to reconstitute nativelike specific nuclear binding sites (acceptor sites) for the oviduct progesterone receptor (PR). These specific nuclear binding sites represent the difference between the binding to the reconstituted NAP and that to pure DNA. The specific fraction of chromatin protein which contains the acceptor activity, fraction CP-3, is very tightly bound to hen DNA in a complex termed nucleoacidic protein (NAP). Removal of the CP-3 fraction from NAP results in a loss of specific PR binding sites. Resins containing chromatin adsorbed to hydroxylapatite are used as a rapid method to isolate the CP-3 fraction. Reconstitution of the CP-3 fraction to DNA by the described method involving a regressing gradient of 6-0 M guanidine hydrochloride (Gdn-HCl) results in a reconstituted NAP which displays specific PR binding sites identical with those in native (undissociated) NAP and whole chromatin. Optimal conditions and potential problems for reconstituting these nucleoproteins are described. Only partially purified receptor preparations were used in these cell-free binding analyses since they have been shown to bind with similar properties and patterns as the nuclear binding in vivo. Therefore, the binding of PR to the reconstituted NAPs was demonstrated to be receptor dependent, saturable, and of high affinity. Further, the pattern of binding to the reconstituted sites mimics those which are observed in vivo. Thus, nonfunctional receptors that cannot translocate and bind to the nuclear acceptor sites in vivo also failed to bind to the acceptor sites on the reconstituted NAPs generated by the acceptor proteins. In contrast, the binding to pure DNA does not reflect these receptor differences in receptor bindings. Specific binding of PR to reconstituted NAP can be reversed by again removing the protein fraction. Moreover, the specific binding can be destroyed by proteases and protected by protease inhibitors, indicating that acceptor activity is proteinaceous in nature. The reconstitution of the activity is both a concentration-dependent and time-dependent process. During the reconstitution, acceptor activity appears to reconstitute on the DNA when the Gdn-HCl concentration reaches 2.0 M. By use of the reconstitution method as an assay for acceptor activity, the activity in the CP-3 fraction was shown by molecular sieve chromatography to elute in a relatively broad molecular weight range between 13 000 and 25 000. The activity also focuses in isoelectric focusing resins with apparent pI's of 5.2 and 6.4.(ABSTRACT TRUNCATED AT 400 WORDS)

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Section: Discussionmentioning

confidence: 99%

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confidence: 84%

Reconstitution of nativelike nuclear acceptor sites of the avian oviduct progesterone receptor: evidence for involvement of specific chromatin proteins and specific DNA sequences

Spelsberg¹,

Gosse

Littlefield

et al. 1984

Biochemistry

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“…Many acidic nuclear proteins have the capacity to bind to DNA. This binding involves noncovalent forces, and the binding capacity is very heterogeneous, involving homologous, heterologous, and double-or single-strand DNA (14,15). In this study, it has been demonstrated that an autoantibody in human disease is directed against a nuclear acidic protein which has the capacity to bind to DNA.…”

Section: Resultsmentioning

confidence: 82%

DNA-binding property of Sm nuclear antigen.

Reyes

Tan

1977

The Journal of Experimental Medicine

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Antibodies to nuclear antigens are detected frequently in the sera of patients with systemic lupus erythematosus (SLE) and related autoimmune diseases. These antibodies have multiple immunochemical specificities and they react with different macromolecular components of cell nuclei (1), among which are nonhistone or nuclear acidic proteins. The first nuclear acidic protein characterized immunochemically with the aid of spontaneously occurring antibodies was called Sm nuclear antigen (2). Subsequently, other nuclear acidic protein antigens have been identified in a similar manner and include a nuclear ribonucleoprotein antigen reactive with sera of patients with SLE and mixed connective tissue disease (3) and other acidic nuclear proteins which are reactive with sera of patients with SjSgren's syndrome, scleroderma, and rheumatoid arthritis (4-6).Many nuclear acidic proteins have been shown to have the property of binding to DNA and to play a role in the control of transcription of DNA to RNA in prokaryotic and eukaryotic systems (7). In the studies reported here, we have shown that tissue extracts containing Sm nuclear antigen have the property of binding to single-strand DNA but not to double-strand DNA. Materials and MethodsSource of Sm Antigen The nuclear origin of the Sm antigen had been established m previous studies (2, 4). The antigen was present in rabbit thymus acetone powder (8) and because this was readily available commercially (Pel-Freez Bio-Animals, Inc., Rogers, Ark.) it was used as the source of Sm antigen. 60 mg of this powder was extracted in each 1 ml of phosphate-buffered saline (0.15 M NaC1, 0.01 M PO4, pH 7.3) by gently stirring the suspension at 4°C for 4 h After centmfugatlon at i0,000 rpm for 10 mm, the supernatant solution contained protein varying from 10 to 15 mg/ml Partml purification of Sm antigen was obtained by salting out between 35 and 60% saturation with ammonium sulfate. This fraction contained all Sm antigenic activity but only onehalf of the total protein and was used as the source material m further studies.Immunologwal Detectwn of Sin Antzgenic Actiwty Sera from patients with SLE containing high tlters of antibody to Sm antigen were used as reagents to detect presence of Sm antigen m isolated fractions of rabbit thymus extract. These sera had no detectable antibody activity to other known nuclear antigen-antibody systems, including DNA, deoxymbonucleoprotein, nuclear mbonucleoprotein, and nuclear antigens A and B reported in Sjogren's syndrome (4). In this sense, these SLE sera were "monospeclfiC" for antibodies to Sm. The immunological assays used were immunodiffusion and inhibition ofpasmve hemagglutmation In the latter test, tanned sheep erythrocytes were coated with rabbit thymus extract according to a procedure that resulted m complexing of Sm antigen to the erythrocytes (9) Presence of Sm antigen in isolated fractions was determined by inhibition of hemagglutmation.

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“…The system described in this paper differs from the aforementioned antigens in that it requires for its immunological specificity the interaction of chromosomal nonhistone proteins with DNA. Based on our experiments with partially purified antigens, the formation of immunologically active complexes involves considerable interaction specificity (Wakabayashi & Hnilica, 1973;Chiu et al, 1974;Wang et al, 1976). Within a given species, the immunological cell or tissue specificity of antisera to dehistonized chromatin preparations depends on the cellular origin of the protein component (Chiu et al, 1974).…”

Section: Discussionmentioning

confidence: 96%

Immunologically specific complexes of chromosomal nonhistone proteins with deoxyribonucleic acid in chicken erythroid nuclei

Krajewska¹,

Briggs²,

Chiu³

et al. 1980

Biochemistry

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Erythroid cell-specific antisera capable of detecting chromosomal nonhistone protein-DNA complexes were obtained by injecting rabbits with dehistonized chicken reticulocyte chromatin. The specific antigenic nonhistone protein-DNA complexes were relatively inaccessible to the antiserum in isolated erythrocyte chromatin. However, isolation of chromatin from cells at earlier stages of erythropoiesis or treatment of isolated erythrocyte chromatin with polyanions or phenylhydrazine provided materials with significantly increased immunological reactivity. The altered activity was caused by changes in conformation occurring at two levels: a specific one, determined by chromosomal nonhistone proteins, and a more general one, determined by histones. Immunological examination of fractionated products obtained from limited nuclease digestion revealed the localization of the antigenic complexes in the nuclease-resistant, large fragments of erythroid chromatin. The nuclease-resistant DNA isolated from the immunologically reactive fragments migrated in gel electrophoresis as a diffuse band of between 1000 and 2000 base pairs. No preferential accumulation of globin-specifying DNA sequences could be found in this nuclease-resistant DNA. The protein fraction containing the immunologically cell-specific complexes in chicken erythrocyte chromatin was glycosylated and moderately acidic (by amino acid anaysis) with an electrophoretically determined Mr of approximately 90000.

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Tissue-specific chromosomal non-histone protein interactions with DNA.

Cited by 42 publications

References 24 publications

Reconstitution of nativelike nuclear acceptor sites of the avian oviduct progesterone receptor: evidence for involvement of specific chromatin proteins and specific DNA sequences

Reconstitution of nativelike nuclear acceptor sites of the avian oviduct progesterone receptor: evidence for involvement of specific chromatin proteins and specific DNA sequences

DNA-binding property of Sm nuclear antigen.

Immunologically specific complexes of chromosomal nonhistone proteins with deoxyribonucleic acid in chicken erythroid nuclei

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