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S Tanuma

Publications and source records attributed to S Tanuma.

At least 91 records · Page 5Linked to original sources
PubMed1986

Occurrence of (ADP-ribose)N glycohydrolase in human erythrocytes.

Chromatography of the cytosol of human erythrocytes on Blue Sepharose, S Sepharose, DEAE-cellulose and single-stranded DNA cellulose yielded a fraction which contained an (ADP-ribose)n glycohydrolase activity. The enzyme hydrolyzed (ADP-ribose)n in a fashion of exo-glycosidase. The native molecular weight of the enzyme was estimated to be 56,000 by gel filtration on Sepharose CL-6B. Its optimum pH was around 7.2. ADP-ribose, cAMP and monovalent salts inhibited the activity. Such characteristics established this glycohydrolase as being a cytosolic glycohydrolase II.

Chromatography, DEAE-Cellulose↗
PubMed1986

Identification of two activities of (ADP-ribose)n glycohydrolase in HeLa S3 cells.

HeLa S3 cells contained two activities (form I and II) that degrade (ADP-ribose)n exo-glycosidically. Form I was extracted from nuclei only by sonication in high ionic strength, while form II was soluble in cytosol. The two active forms differed in chromatographic behaviors, in their Km values for (ADP-ribose)n, and in their pH and salt requirements for optimal activity, although both forms exhibited properties characteristic of (ADP-ribose)n glycohydrolase such as requirement of sulfhydryl compounds and sensitivity of ADP-ribose and cAMP. Form I and II had apparent molecular weights of 72,000 and 53,000, respectively, as determined by gel filtration on Sepharose CL-6B.

Adenosine Diphosphate Ribose↗
PubMed1986

Purification and properties of an (ADP-ribose)n glycohydrolase from guinea pig liver nuclei.

An (ADP-ribose)n glycohydrolase has been purified more than 3,000-fold from guinea pig liver nuclei with an 18% yield. The glycohydrolase activity present in the nuclei was solubilized only by sonication at high ionic strength and purified by sequential chromatographic steps on phosphocellulose, DEAE-cellulose, Blue Sepharose, and single-stranded DNA cellulose. The purified protein exhibited one predominant protein band on sodium dodecyl sulfate-polyacrylamide gels with an estimated molecular weight of 75,500. On Sephadex G-100 gel filtration, single coincident peaks of (ADP-ribose)n glycohydrolase activity and protein with a molecular weight value of 72,000 were observed. The Km value for (ADP-ribose)n and the maximal velocity of the highly purified glycohydrolase were 2.3 microM and 36 mumol of ADP-ribose released from (ADP-ribose)n . min-1 . mg protein-1, respectively. Hydrolysis of (ADP-ribose)n by the enzyme was exoglycosidic in nature. The optimum pH for the enzyme activity was apparent at 6.8-7.0. Sulfhydryl compounds and monovalent cations were required for the maximal activity. The enzyme was sensitive to Ca2+ but not to Mg2+. The enzyme activity was inhibited by ADP-ribose, cyclic AMP (adenosine 3':5'-monophosphate) and diadenosine 5',5'''-p1,p4-tetraphosphate. Denatured DNA and histones were inhibitory, but native DNA and its histone complex were not inhibitory. Our data indicate that the glycohydrolase is present only as a minor protein in nuclei, being present in perhaps about 50,000 molecules/nucleus.

Animals↗
PubMed1986

Acceptor proteins for (ADP-ribose)n in the HeLa S3 cell cycle.

The acceptor proteins for (ADP-ribose)n were investigated by using nuclei or chromosomes isolated from specific phases of the cell cycle of HeLa S3 cells. Analysis of HMG proteins and histone H1 by acetic acid/urea polyacrylamide gel electrophoresis demonstrated that the (ADP-ribosyl)n-ation of HMG 14 and 17 and histone H1 increased by 12- and 5-fold, respectively, in the metaphase chromosomes as compared with that in the G1 phase cell nuclei. The degree of (ADP-ribosyl)n-ation of these proteins in the S phase cell nuclei was as low as that in G1 phase cell nuclei. In the G2 phase cell nuclei, the degrees of (ADP-ribosyl)n-ation of HMG 14 and 17 and histone H1 were about 5- and 2-fold greater, respectively, as compared with that in the G1 phase cell nuclei. The (ADP-ribosyl)n-ation of HMG 1 and 2 was constant through the cell cycle except for a slight decrease in the S phase. The data may imply that the (ADP-ribosyl)n-ation of HMG 14 and 17 and histone H1 is linked to chromatin structural changes in mitosis.

Cell Cycle↗
PubMed1985

Comparison of ADP-ribosylation of chromosomal proteins between intact and broken cells.

ADP-ribosylation of nonhistone high mobility group (HMG) proteins and histone H1 in intact cells was markedly different from that in the broken cell systems using isolated nuclei and chromatin. (i) The amounts of (ADP-ribose)n on these proteins in intact cells were less 10(-2) - 10(-3) of those in the broken cell systems. (ii) The modified protein molecules in intact cells were about 2 orders of magnitude less extent than those in the broken cells. (iii) The (ADP-ribose)n chains synthesized in intact cells, nuclei and chromatin were mainly mono, oligo and poly, respectively. (iv) The principal acceptor molecules in intact cells were concentrated in HMG 1, 2, 14 and 17, and histone H1, whereas the four HMG proteins were minor acceptors in nuclei, and HMG 1 and 2 and histone H1 were major acceptors in chromatin.

Adenosine Diphosphate Ribose↗
PubMed1985

Endogenous ADP ribosylation of high mobility group proteins 1 and 2 and histone H1 following DNA damage in intact cells.

Endogenous ADP ribosylation of nonhistone high-mobility group (HMG) proteins and histone H1 was studied in cultured mouse mammary tumor cells following treatment with N-methyl-N'-nitro-N-nitrosoguanidine (MNNG). MNNG treatment of cells caused a rapid and transient increase in ADP ribosylation of histone H1 and HMG 1 and 2, whereas (ADP-ribose)n on HMG 14 and 17 was not affected. 3-Aminobenzamide, an inhibitor of (ADP-ribose)n synthetase, prevented the increase in ADP ribosylation of histone H1 and HMG 1 and 2. This inhibitor enhanced the cell-killing effect of MNNG, but had no significant effect on the removal of methylated purines. The preferential increase in ADP ribosylation of HMG 1 and 2 and histone H1 may be necessary for cell recovery from DNA damage.

Adenosine Diphosphate Ribose↗
PubMed1984

Maturation of human promyelocytic leukemia cells induced by nicotinamide: evidence of a regulatory role for ADP-ribosylation of chromosomal proteins.

We have studied the role of ADP-ribosylation of chromosomal proteins in the regulation of myeloid cell maturation using the HL-60 cell line as a model. Nuclei isolated from this human promyelocytic leukemia cell line contained (ADP-ribose)n synthetase activity, whereas little or no enzymatic activity was detectable in normal human blood neutrophils. Furthermore, the activity of (ADP-ribose)n synthetase was decreased in HL-60 cells when they were induced to mature with retinoic acid (RA). To determine whether reduced (ADP-ribose)n synthetase activity is simply a result of induced maturation or whether it is a necessary precedent event for the maturation process, we evaluated the effects of nicotinamide (NAm) and its methyl derivative, N'-methylnicotinamide (N'-Met-NAm), agents which decrease ADP-ribosylation. Treatment of HL-60 cells with these drugs caused the cells to undergo maturation and to acquire certain of the morphologic, functional, and biochemical characteristics of normal neutrophils. N'-Met-NAm was more potent than NAm in inducing maturation; at a concentration of 0.8 mM, it caused greater than 80% of the cells to mature, whereas a tenfold greater concentration of NAm was required to induce a similar degree of maturation. NAm and N'-Met-NAm also potentiated the maturation of HL-60 cells induced by RA. Exposure of cells to noninducing concentrations of these compounds caused a leftward shift in the dose-response curve for RA; maturation was observed at 10(-11) M RA in the presence of either 2 mM NAm or 0.2 mM N'-Met-NAm while 10(-9) M RA was required to induce maturation in their absence. A leftward shift in the dose response curve for maturation in the presence of low doses of NAm or N'-Met-NAm did not occur with another inducer, dimethyl formamide (DMF). Two enzymes, NAD glycohydrolase and tissue transglutaminase, that are abundant in macrophages, were induced by RA but not by NAm. N'-Met-NAm decreased by about 75% the amount of endogenous (ADP-ribose)n in a selected fraction of chromosomal proteins which included histone H1 and the nonhistone high mobility group proteins. The results of this study support the concept that ADP-ribosylation of chromosomal proteins influences the regulation of human myeloid cell maturation.

Acyltransferases↗
PubMed1983

ADP-ribosylation of chromosomal proteins and mouse mammary tumor virus gene expression. Glucocorticoids rapidly decrease endogenous ADP-ribosylation of nonhistone high mobility group 14 and 17 proteins.

The relationship between endogenous ADP-ribosylation of chromosomal proteins and glucocorticoid-regulated mouse mammary tumor virus gene expression was investigated in cultured mouse mammary tumor cells. It was observed that glucocorticoids quickly decreased endogenous (ADP-ribose)n on the nonhistone high mobility group (HMG) 14 and 17 proteins. The half-time for this loss was 8 and 17 min, respectively, for the two proteins. (ADP-ribose)n on HMG 1 and 2 and on histone H1 was less susceptible to hydrolysis during glucocorticoid treatment. The rapid loss of (ADP-ribose)n from HMG 14 and 17 occurred in the same time frame as the induction of mouse mammary tumor virus RNA synthesis by glucocorticoids in these cells (Young, H. A., Shih, T. Y., Scolnick, E. M., and Parks, W. P. (1977) J. Virol. 21, 139-149). 3-Amino-benzamide, a specific inhibitor of (ADP-ribose)n synthetase, increased mouse mammary tumor virus RNA levels with an accompanying decrease in endogenous ADP-ribosylation of HMG 14 and 17. These results show that a decrease in endogenous ADP-ribosylation of HMG 14 and 17 is a consequence of glucocorticoid action and suggest that loss of (ADP-ribose)n from these proteins may be an important event in mouse mammary tumor virus gene expression.

Adenosine Diphosphate Ribose↗
PubMed1983

ADP-ribosylation of nonhistone high mobility group proteins in intact cells.

The ADP-ribosylation of nonhistone, high mobility group (HMG) proteins in intact cultured cells was investigated. Radioactively labeled adenosine was used as a precursor to detect (ADP-ribose)n on protein. A protein fraction enriched in HMG proteins and histone H1 was separated from RNA and DNA by CsCl density gradient centrifugation, 5% perchloric acid extraction, and CM-Sephadex C-50 column chromatography. Poly- and mono(ADP-ribose) were recovered following alkaline hydrolysis, and 5'-AMP and (2'-(5"-phosphoribosyl)-5'-AMP) were produced by phosphodiesterase treatment, indicating that the protein-bound radioactive material was (ADP-ribose)n. An average chain length of 1.5 to 1.8 was determined. Analysis of proteins by sodium dodecyl sulfate and acetic acid/urea polyacrylamide gel electrophoresis demonstrated that HMG 1, 2, 14, and 17 as well as histone H1 contained (ADP-ribose)n. Treatment of cells with 3-aminobenzamide, an inhibitor of (ADP-ribose)n synthetase, decreased endogenous ADP-ribosylation in both types of chromosomal proteins but that of HMG 14 and 17 was affected more.

Adenosine Diphosphate Ribose↗
PubMed1983

Purification and characterization of two forms of DNA polymerase alpha from HeLa cell nuclei.

Isolated nuclei contained two active forms of DNA polymerase alpha (form I and form II). Form II was extracted from nuclei by KCl at concentrations lower than 0.18 M. Above 0.18 M selective extraction of form I was observed. The purified two active forms differed in chromatographic and electrophoretic behaviors, in their salt requirement for optimal activity, and in preference of template-primers, although both forms exhibited properties characteristic of DNA polymerase alpha such as sensitivity of N-ethylmaleimide, 1-beta-D-arabino-furanosylcytosine triphosphate, and aphidicolin. Marked difference between the two forms was preference of template-primer that form I was more active with poly(dT).(rA)10 than poly(dA).(dT)12 whereas form II exhibited higher activity with poly(dA).(dT)12 than poly(dT).(rA)10. Possible roles of two forms of DNA polymerase alpha in the processes of DNA replication will be discussed.

Cell Nucleus↗
PubMed1982

Poly(ADP-ribosyl)ation of chromosomal proteins in the HeLa S3 cell cycle.

Change in the level of poly(ADP-ribosyl)ation during the cell cycle of HeLa S3 cells was examined in three different in vitro systems: isolated nuclei, chromatin, and poly(ADP-ribose) polymerase extracted from isolated nuclei, and by the direct immunofluorescent antibody technique in situ. In isolated nuclei, after release from thymidine-hydroxyurea double block, the level of poly(ADP-ribosyl)ation was low in the S phase and increased in the late G2 phase with a peak in the M phase. In chromatin, the level began to increase in the mid-late S phase, was maintained at a high level throughout the G2 phase, decreased in the M phase, and became basal during the G1 phase. The activity of poly(ADP-ribose) polymerase extracted from isolated nuclei was relatively high throughout the cell cycle and was highest in the G2 phase. Similar changes during the HeLa S3 cell cycle were observed in the three systems after release from Colcemid block. In synchronously growing HeLa S3 cells in situ, nuclei in the G2-M phase were stained strongest for poly(ADP-ribose) by the immunofluorescent staining. These results were consistent with the peak of poly(ADP-ribosyl)ation in the G2-M phase observed in the nuclear system in vitro.

Cell Cycle↗
PubMed1982

Absence of response of chick embryonic limb to the growth stimulatory effect of parathyroid hormone in vitro after exposure to 6-aminonicotinamide in ovo.

The biochemical effect of a nicotinamide analogue, 6-aminonicotinamide (6-AN), on developing chick embryonic femur was studied. Growth of femur from 9-day-old embryos that had been exposed to 6-AN for 5 days in ovo was not stimulated by PTH in an in vitro culture system. PTH caused a much smaller increase in the cyclic-AMP content in 6-AN-treated femur than in control femur. However, dibutyryl cyclic-AMP stimulated growth of 6-AN-treated femur. A defect in response of 6-AN-treated femur to the growth stimulating action of PTH may explain the production of micromelia by 6-AN.

6-Aminonicotinamide↗
PubMed1981

Immunofluorescent staining of poly(ADP-ribose) in situ in HeLa cell chromosomes in the M phase.

Randomly and synchronously growing HeLa cells were tested for poly(ADP-ribose) by direct and indirect immunofluorescent antibody techniques. Fluorescence of poly(ADP-ribose) was seen only in the nuclei of intact cells when the direct immunofluorescent antibody technique was used but in both the nuclei and cytoplasm when the indirect immunofluorescent antibody technique was used; fluorescence in the cytoplasm was nonspecific. When randomly or synchronously growing HeLa cells were fixed in acetone and treated with DNase I before incubation with fluorescein-labeled antibody, intense fluorescence was observed only in the nuclei when the direct immunofluorescent staining technique was used. Addition of 3-aminobenzamide, a potent inhibitor of poly(ADP-ribose) polymerase, with the DNase I completely abolished the fluorescence in the nuclei of synchronously and randomly growing HeLa cells, except in M-phase nuclei. These results suggest that poly(ADP-ribose) can be synthesized even in the nuclei of acetone-fixed HeLa cells from "endogenous NAD+" during incubation with fluorescent antibody and also that the fluorescence of chromosomes of HeLa cells in the M phase is, in fact, due to the in situ presence of poly(ADP-ribose), not to poly(ADP-ribose) synthesized during incubation with antibody.

Chromosomes, Human↗