Enzyme-bound early product of purified poly(ADP-ribose) polymerase

Yoshihara, Koichiro; Hashida, T; Yoshihara, H; Tanaka, Yasuharu; Ohgushi, Hajime

doi:10.1016/0006-291x(77)91431-0

Cited by 126 publications

(35 citation statements)

References 28 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The catalytically active species may well be an oligomer of this polypeptide chain. The molecular weight of the monomer agrees fairly well with those reported for polymerase from some other tissues: 130000 for the bovine thymus enzyme [20] and 120000 for the calf thymus enzyme [22], while it is significantly different from the values of 50000 reported for the rat liver enzyme [I91 and 63 500 for the pig thymus enzyme 1211. The present study is, however, the only one in which the major protein component of the preparation has been shown to be responsible for the synthesis of poly(ADP-ribose).…”

Section: Discussionsupporting

confidence: 82%

Poly (ADP‐ribose) Polymerase from Ehrlich Ascites Tumor Cells

Kristensen

Holtlund

1978

European Journal of Biochemistry

View full text Add to dashboard Cite

Poly(ADP-ribose) polymerase from Ehrlich ascites tumor cells, partially purified by chromatography on DNA-agarose, was obtained as a more than 80 ' %, homogeneous preparation by isoelectric focusing in a sucrose gradient. The polymerase activity was shown to be associated with the major protein in the preparation. Results obtained by electrophoresis in the presence of sodium dodecylsulfate indicated that poly(ADP-ribose) polymerase consists of a polypeptide chain with a molecular weight of 130 000. Ultracentrifugation at non-denaturating conditions indicated that the active enzyme may be an oligomeric form of this polypeptide chain. The isoelectric point of the polymerase was 9.40. The effects of various additions to the assay mixture on the synthesis of poly(ADPribose) as well as some kinetic data, are given. It is shown that poly(ADP-ribose) is a highly efficient inhibitor of its own synthesis, and results are presented which suggest that the well-known stimulatory effect of DNA on the synthesis is due to reduction of this inhibitory effect of the product.Eukaryotic cell nuclei contain an enzyme which incorporates the ADP-ribose moiety of NAD into poly(ADP-ribose) [l -31. Poly(ADP-ribose) of various sizes, with chain length varying from 1-40, is bound to nuclear proteins, both histones and non-histones [4,5]. Although the biological role of the polymer is not yet understood, its involvement in the control of DNA synthesis [6][7][8], DNA repair [9-121, cell proliferation [13-151 and cell differentiation has been reported.Recently, several procedures for the isolation of highly purified poly(ADP4bose) polymerase from different sources have been reported [I9 -221. The availability of purified polymerase may be of great importance, both in elucidating the biological role of poly(ADP-ribose) and in other kinds of studies. It is, for instance, not known how the cell regulates the degree of ADP-ribosylation of the different acceptor proteins and whether there exist several polymerases with different specificities towards the acceptor proteins. Also, polymerase purification will make it possible to determine whether the higher rate of synthesis of poly(ADP-ribose) reported in transformed cells [23] is due to higher polymerase concentrations or to the stances, has the major protein present after purification been shown to be responsible for the polymer synthesis. In the present work we will describe a relatively fast and simple method for obtaining an almost homogeneous preparation of poly(ADP-ribose) polymerase from Ehrlich ascites tumor cells. The major protein present is shown to be able to synthesize poly-(ADP-ribose). Some properties of the polymerase are given, as well as a possible explanation of the wellknown DNA requirement of the polymer synthesis [24,25].

show abstract

Section: Discussionsupporting

confidence: 82%

Poly (ADP‐ribose) Polymerase from Ehrlich Ascites Tumor Cells

Kristensen

Holtlund

1978

European Journal of Biochemistry

View full text Add to dashboard Cite

show abstract

“…In contrast, several groups showed that highly purified PARP-1 was unable to covalently poly-ADP-ribosylate highly purified histones (298,451). The only observed acceptor protein for poly-ADP-ribose was PARP-1.…”

Section: Pargsmentioning

confidence: 99%

Nuclear ADP-Ribosylation Reactions in Mammalian Cells: Where Are We Today and Where Are We Going?

Hassa

Haenni

Elser

et al. 2006

Microbiol Mol Biol Rev

631

644

View full text Add to dashboard Cite

SUMMARY Since poly-ADP ribose was discovered over 40 years ago, there has been significant progress in research into the biology of mono- and poly-ADP-ribosylation reactions. During the last decade, it became clear that ADP-ribosylation reactions play important roles in a wide range of physiological and pathophysiological processes, including inter- and intracellular signaling, transcriptional regulation, DNA repair pathways and maintenance of genomic stability, telomere dynamics, cell differentiation and proliferation, and necrosis and apoptosis. ADP-ribosylation reactions are phylogenetically ancient and can be classified into four major groups: mono-ADP-ribosylation, poly-ADP-ribosylation, ADP-ribose cyclization, and formation of O-acetyl-ADP-ribose. In the human genome, more than 30 different genes coding for enzymes associated with distinct ADP-ribosylation activities have been identified. This review highlights the recent advances in the rapidly growing field of nuclear mono-ADP-ribosylation and poly-ADP-ribosylation reactions and the distinct ADP-ribosylating enzyme families involved in these processes, including the proposed family of novel poly-ADP-ribose polymerase-like mono-ADP-ribose transferases and the potential mono-ADP-ribosylation activities of the sirtuin family of NAD+-dependent histone deacetylases. A special focus is placed on the known roles of distinct mono- and poly-ADP-ribosylation reactions in physiological processes, such as mitosis, cellular differentiation and proliferation, telomere dynamics, and aging, as well as “programmed necrosis” (i.e., high-mobility-group protein B1 release) and apoptosis (i.e., apoptosis-inducing factor shuttling). The proposed molecular mechanisms involved in these processes, such as signaling, chromatin modification (i.e., “histone code”), and remodeling of chromatin structure (i.e., DNA damage response, transcriptional regulation, and insulator function), are described. A potential cross talk between nuclear ADP-ribosylation processes and other NAD+-dependent pathways is discussed.

show abstract

“…5B, slot 1). The (23) have reported that poly(ADP-ribose) polymerase isolated from calf thymus nuclei has a Mr of 130,000. They showed that in an in vitro reaction, the enzyme was capable of self-modification.…”

Section: Preferential Polynucleosome Cleavage Sitesmentioning

confidence: 99%

Nucleosome periodicity in HeLa cell chromatin as probed by micrococcal nuclease.

Butt¹,

Jump²,

Smulson³

1979

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

View full text Add to dashboard Cite

When HeLa cell nuclei were treated with micrococcal nuclease (nucleate 3-oligonucleotidohydrolase, EC 3.1.4.7), lysed, and centrifuged, the supernatant from early digests contained two predominant classes of polynucleosomes of repeat size 8N and 16N. With increasing digestion time, the 16N polynucleosome appeared to be cleaved to the 8N species and finally to the basic subunit of chromatin. The size of the polynucleosomes has been determined by DNA analysis and on polyacrylamide electrophoretic gels of native chromatin particles. Thel6Npolynucleosome appears to be a unique higher ordered structural component of HeLa cell chromatin. Our recent report, showing that the nuclear protein-modifying enzyme poly(ADP-ribose) polymerase increases in specific activity progressively with increasing nucleosome repeat size up to 8-1ON, has been extended in the present study. Activity was also elevated in the polynucleosomes of the 16N structure preferentially cleaved by micrococcal nuclease, although specific activity of the enzyme was highest in octanucleosomes. Acceptors for poly(ADP-ribose) have also been determined in these particles.Despite recent advancement in our understanding of the structure of the nucleosome core particle, similar information concerning the higher order structure of chromatin remains largely undetermined (1)(2)(3). At low ionic strength the nucleofilament with diameter of 100 A is a single chain of nucleosomes (4-7). In the presence of monovalent and divalent cations, the diameter of the nucleofilament is increased to 250400 A (4, 5, 7). Based on electron micrographic data, Finch and Klug (4) postulated a solenoid model of chromatin.Kinetic studies performed on the action of micrococcal nuclease (nucleate 3'-oligonucleotidohydrolase, EC 3.1.4.7) on interphase chromatin and metaphase chromosomes (8-10) have supported the theory of random cleavage by the enzyme at uniformly susceptible sites on linker regions of polynucleosomes. However, it has also been shown that newly replicated chromatin is relatively more susceptible to the action of micrococcal nuclease (11).Using electron microscopic studies and sedimentation analysis on the chromatin produced after the digestion of lymphocyte nuclei by micrococcal nuclease, Hozier et al. (7) have postulated that chromatin is organized into superbeads containing six to ten nucleosomes. However, ambiguity about these structures still remains since Hozier et al. (7) were unable to isolate and resolve them on sucrose gradients. In kinetic studies performed in the work below, it has clearly been shown that micrococcal nuclease cleaves HeLa chromatin at a periodicity of eight nucleosomes. In addition to sedimentation analysis, we have been able to characterize these structures by (i) native chromatin gels, (ai) determination of the size of the DNA, and (Mi) the presence of a full complement of nuclear histones.These observations on nucleosome periodicity stemmed directly from our previous observation that the specific activity of a chromatin-bound enzyme, ...

show abstract

Enzyme-bound early product of purified poly(ADP-ribose) polymerase

Cited by 126 publications

References 28 publications

Poly (ADP‐ribose) Polymerase from Ehrlich Ascites Tumor Cells

Poly (ADP‐ribose) Polymerase from Ehrlich Ascites Tumor Cells

Nuclear ADP-Ribosylation Reactions in Mammalian Cells: Where Are We Today and Where Are We Going?

Nucleosome periodicity in HeLa cell chromatin as probed by micrococcal nuclease.

Contact Info

Product

Resources

About