Noncovalent Interaction between Poly(ADP-Ribose) and Cellular Proteins: An Application of a Poly(ADP-Ribose) Western Blotting Method to Detect Poly(ADP-Ribose) Binding on Protein-Blotted Filter

Nozaki, Tadashige; Masutani, Mitsuko; Akagawa, Takumi; Sugimura, T.; Esumi, Hiroyasu

doi:10.1006/bbrc.1994.1007

Cited by 22 publications

(12 citation statements)

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“…Due to the fact that the nuclear matrix is devoid of histone proteins, we conclude that some of the nuclear matrix proteins also interact ionic ally with free poly(ADP-ribose) and inhibit PARG. Our conclusion is compatible with the previously reported non-covalent interactions of poly(ADP-ribose) with histone proteins as examined by: (i) a phenol/partitioning assay [11]; (ii) a poly(ADP-ribose )-westem blotting approach [18], and a photoaffinity labeling procedure [19] . In our poly(ADP-ribose) preparations, the lack of monomeric ADPribose allowed us to simultaneously monitor PARG activity as well as non-covalent interactions of long polymers with chromatin protein(s) .…”

Section: Discussionsupporting

confidence: 91%

“…In our poly(ADP-ribose) preparations, the lack of monomeric ADPribose allowed us to simultaneously monitor PARG activity as well as non-covalent interactions of long polymers with chromatin protein(s) . Nozaki et at., [18] showed that the polypeptides of a HeLa cell nuclear extract strongly interacted non-covalently with ADP-ribose chains larger that 20 residues. Although histone proteins were the most prominent proteins identified, it should be noted that some of the most efficient polypeptides that blot radiolabeled poly(ADP-ribose) electrophoretic ally migrate in the molecular weight range of the nuclear matrix lamins.…”

Section: Discussionmentioning

confidence: 99%

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Measurement of poly(ADP-ribose) glycohydrolase activity by high resolution polyacrylamide gel electrophoresis: Specific inhibition by histones and nuclear matrix proteins

Pacheco–Rodriguez

Alvarez-Gonzalez

1999

ADP-Ribosylation Reactions: From Bacterial Pathogenesis to Cancer

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We have developed a novel enzyme assay that allows the simultaneous determination of noncovalent interactions of poly(ADP-ribose) with nuclear proteins as well as poly(ADP-ribose) glycohydrolase (PARG) activity by high resolution polyacrylamide gel electrophoresis. ADP-ribose chains between 2 and 70 residues in size were enzymatically synthesized with pure poly(ADP-ribose) polymerase (PARP) and were purified by affinity chromatography on a boronate resin following alkaline release from protein. This preparation of polymers of ADP-ribose was used as the enzyme substrate for purified PARG. We also obtained the nuclear matrix fraction from rat liver nuclei and measured the enzyme activity of purified PARG in the presence or absence of either histone proteins or nuclear matrix proteins. Both resulted in a marked inhibition of PARG activity as determined by the decrease in the formation of monomeric ADP-ribose. The inhibition of PARG was presumably due to the non-covalent interactions of these proteins with free ADP-ribose polymers. Thus, the presence of histone and nuclear matrix proteins should be taken into consideration when measuring PARG activity. (Mol Cell Biochem 193: 13-18, 1999)

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Section: Discussionsupporting

confidence: 91%

Section: Discussionmentioning

confidence: 99%

Measurement of poly(ADP-ribose) glycohydrolase activity by high resolution polyacrylamide gel electrophoresis: Specific inhibition by histones and nuclear matrix proteins

Pacheco–Rodriguez

Alvarez-Gonzalez

1999

ADP-Ribosylation Reactions: From Bacterial Pathogenesis to Cancer

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“…The membranes were incubated with the primary and the secondary antibodies in Tris-buffered saline with 0.05% Tween 20 for 60 min and then were washed in the same medium. The primary antibodies for these experiments included mouse monoclonal antibody for poly(ADP-ribose) (23,40) and rabbit polyclonal antibody for ␥-tubulin. The bound primary antibody was detected with alkaline phosphatase-conjugated secondary antibody and visualized with NBT/ BCIP (Roche).…”

Section: Cells Parp-1mentioning

confidence: 99%

“…Many proteins that are poly-(ADP-ribosyl)ated by PARP-1 have been identified, including PARP-1 itself (43), histones (26), lamins (1), topoisomerases (25), DNA polymerases (44,70), c-Fos (2), and p53 tumor suppressor protein (68). Since the attachment of the negatively charged polymer changes the properties of the acceptor protein (40,46), PARP-1 could be involved in a variety of cellular events, including modulation of chromatin structure, DNA synthesis, DNA repair, gene transcription, and cell cycle reg-ulation (13). In particular, the studies with PARP inhibitors have shown that PARP-1 plays an important role in maintenance of genome integrity (10,34,35).…”

mentioning

confidence: 99%

Involvement of Poly(ADP-Ribose) Polymerase 1 and Poly(ADP-Ribosyl)ation in Regulation of Centrosome Function

Kanai

Tong

Sugihara

et al. 2003

Molecular and Cellular Biology

136

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The regulatory mechanism of centrosome function is crucial to the accurate transmission of chromosomes to the daughter cells in mitosis. Recent findings on the posttranslational modifications of many centrosomal proteins led us to speculate that these modifications might be involved in centrosome behavior. Poly(ADPribose) polymerase 1 (PARP-1) catalyzes poly(ADP-ribosyl)ation to various proteins. We show here that PARP-1 localizes to centrosomes and catalyzes poly(ADP-ribosyl)ation of centrosomal proteins. Moreover, centrosome hyperamplification is frequently observed with PARP inhibitor, as well as in PARP-1-null cells. Thus, it is possible that chromosomal instability known in PARP-1-null cells can be attributed to the centrosomal dysfunction. P53 tumor suppressor protein has been also shown to be localized at centrosomes and to be involved in the regulation of centrosome duplication and monitoring of the chromosomal stability. We found that centrosomal p53 is poly(ADP-ribosyl)ated in vivo and centrosomal PARP-1 directly catalyzes poly(ADPribosyl)ation of p53 in vitro. These results indicate that PARP-1 and PARP-1-mediated poly(ADP-ribosyl)ation of centrosomal proteins are involved in the regulation of centrosome function.

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“…Although short ADP-ribose oligomers, as well as long polymers, have been shown to bind noncovalently to histones (Panzeter et al, 1992), polymers longer than 20 ADP-ribose units exhibit higher binding affinity (Panzeter et al, 1992;Nozaki et al, 1994). Therefore it seemed important to investigate whether or not such poly-ADPR molecules were synthesized in rat testes under physiologic conditions.…”

Section: Analysis Of Adp-ribose Polymers Synthesized In Vivo In Rat Tmentioning

confidence: 98%

Noncovalent Binding of Poly(ADP-Ribose) to Nuclear Matrix Proteins: Developmental Changes and Tissue Specificity

Malanga

Farina²

2000

Biological Chemistry

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Poly(ADP-ribose) is a nuclear polynucleotide involved in the regulation of chromatin functions via covalent and/or noncovalent modification of nuclear proteins. Using a binding assay on protein blots, we searched for poly(ADP-ribose) binding proteins in nuclear matrices from testes of differently aged rats as well as from various adult rat tissues (brain, liver, spleen). We found that nuclear matrix proteins represent a significant subset of the nuclear proteins that can establish noncovalent interactions with poly(ADP-ribose). The profiles of poly(ADP-ribose) binding nuclear matrix proteins appeared to be tissue-specific and changed during postnatal development in the testis. The isolation and analysis of endogenous poly-(ADP-ribose) from rat testes showed that the ADP-ribose polymers that bind nuclear matrix proteins in vitro are also present under physiologic conditions in vivo. These results further substantiate the possibility that poly(ADP-ribose) may affect chromatin functions through noncovalent interaction with specific protein targets, including nuclear matrix components.

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Noncovalent Interaction between Poly(ADP-Ribose) and Cellular Proteins: An Application of a Poly(ADP-Ribose) Western Blotting Method to Detect Poly(ADP-Ribose) Binding on Protein-Blotted Filter

Cited by 22 publications

References 0 publications

Measurement of poly(ADP-ribose) glycohydrolase activity by high resolution polyacrylamide gel electrophoresis: Specific inhibition by histones and nuclear matrix proteins

Measurement of poly(ADP-ribose) glycohydrolase activity by high resolution polyacrylamide gel electrophoresis: Specific inhibition by histones and nuclear matrix proteins

Involvement of Poly(ADP-Ribose) Polymerase 1 and Poly(ADP-Ribosyl)ation in Regulation of Centrosome Function

Noncovalent Binding of Poly(ADP-Ribose) to Nuclear Matrix Proteins: Developmental Changes and Tissue Specificity

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