Purification and Characterization of the Ca2+ Plus Mg2+-Dependent Endodeoxyribonuclease from Calf Thymus Chromatin1

Nakamura, Masashi; Sakaki, Yoshiyuki; Watanabe, Nobuko; Takagi, Yukio

doi:10.1093/oxfordjournals.jbchem.a133175

Cited by 68 publications

(17 citation statements)

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“…A similar Ca2+/Mg2+-dependent endonuclease activity has also been described in the nuclei of rat liver cells (20,21), calf thymocytes (27), and mouse gut epithelial cells (23 (28) and in activation of the endogenous endonuclease in isolated nuclei (20,21,23,27); (ii) Zn2+ inhibits DNA fragmentation in target cells (Table 3) and Zn2+ inhibits the endonuclease activity in thymocytes incubated with glucocorticoids and in isolated thymocyte nuclei incubated with Ca2+ (16); and (iii) DNA fragments found in target cells consist of multiples of approximately 200-base-pair subunits (Fig. 2) and similarly sized fragments are found in thymocytes incubated with glucocorticoids (15,26) and in isolated nuclei of several cell types incubated with Ca2+ and Mg2+ (20,21,23,27).…”

Section: Discussionsupporting

confidence: 67%

Endogenous endonuclease-induced DNA fragmentation: an early event in cell-mediated cytolysis.

Duke¹,

Chervenak²,

Cohen³

1983

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

530

216

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Within minutes of exposure of target cells to cytotoxic T lymphocytes, their nuclear DNA begins to be fragmented. This phenomenon precedes 5"Cr release by at least an hour. DNA fragmentation occurs only when appropriately sensitized cytotoxic T cells are used and is not merely a result of cell death because killing of target cells by heating, freeze/thawing, or lysing with antibody and complement did not yield DNA fragments. Agarose gel electrophoresis of target cell DNA showed discrete multiples of an approximately 200-base-pair subunit, suggesting that fragmentation was the result of activation of a specific endonuclease. A similar pattern of DNA fragments is observed during glucocorticoid-induced killing of mouse thymocytes. The endonuclease in that case is inhibited by zinc ions, and we find that Zn2+ also inhibits DNA fragmentation and 51Cr release induced by cytotoxic T cells, suggesting a final common biochemical pathway for both types of cell death.

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Section: Discussionsupporting

confidence: 67%

Endogenous endonuclease-induced DNA fragmentation: an early event in cell-mediated cytolysis.

Duke¹,

Chervenak²,

Cohen³

1983

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

530

216

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“…While estimates of free cytoplasmic Ca2+ ions have been made in Friend cells (0.14 ,uM) (6) and other tissues (0.11-0.97 ,uM) (33), the level of free intranuclear Ca2+ is not known. Purified Ca2+,Mg2+-dependent endonucleases from calf thymus (34) and bull semen (35) require 0.5-1 mM for optimal activity, but this finding is difficult to assess in terms of what concentration might be required for activity in the cell. Still, the finding that Ca2+ ion influx and endonuclease activity occur during a similar early period of erythroid differentiation suggests that the two events may be related.…”

Section: Discussionmentioning

confidence: 99%

Induction of a Ca2+, Mg2+-dependent endonuclease activity during the early stages of murine erythroleukemic cell differentiation.

McMahon¹,

Alsina²,

Levy³

1984

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

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A Ca2+,Mg2+-dependent endonuclease activity was detected in erythroleukemic cells undergoing differentiation in vitro in response to induction by dimethyl sulfoxide (Me2SO) or hexamethylene-bis-acetamide (HMBA). The endonuclease activity was demonstrated in isolated nuclei within 6 hr after the addition of inducer, reached maximum levels between 24 and 48 hr, and returned to control levels within 72 hr. The activity caused single strand breaks in high molecular weight native DNA, which could be labeled at exposed 3'-OH termini with Escherichia coli DNA polymerase I and radiolabeled nucleotides. Alkaline elution studies revealed DNA fragmentation that appeared coincident with the presence of the endonuclease activity. The detection and levels of single strand DNA breakage correlated with induction of terminal differentiation by Me2SO or HMBA. Induction of the endonuclease activity was reversible: depletion of Me2SO from the growth medium after treatment for 6 and 18 hr led to a rapid decrease in the level of activity. Removal of the inducer prevented terminal differentiation, a finding that strongly suggests the endonuclease activity is present during the precommitment phase of differentiation. DNA fragmentation was not observed in cells incubated with hemin, which has been shown previously to increase the cytoplasmic level of globin mRNA without causing commitment to terminal maturation. Me2SO did not induce the endonuclease activity or DNA fragmentation in an uninducible Friend cell line.

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“…Group D showed a significantly higher survival compared with group B (P < 0.01; Scheffe's test). endonucleases are activated in the presence of manganese [49][50][51]. It is of interest that human serum includes 33 Ϯ 11 mM of manganese [52], and this concentration of manganese shows an inhibitable effect on neutrophil apoptosis, as seen in Fig.…”

Section: Figmentioning

confidence: 99%

Inhibition of Neutrophil Apoptosis by Antioxidants in Culture Medium

Oishi

Machida

1997

Scand J Immunol

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Oishi K, Machida K. Inhibition of Neutrophil Apoptosis by Antioxidants in Culture Medium. Scand J Immunol 1997;45:21-27 Neutrophil apoptosis is an important mechanism that has implications for understanding the life span and toxic potentials of neutrophils at inflamed sights. In this study the authors examined the possibility that reactive oxygen species (ROS) released by neutrophils can regulate neutrophil survival. Cu,Zn-superoxide dismutase (Cu,Zn-SOD), Mn-SOD, and catalase in culture media were significantly effective in delaying the spontaneous apoptosis, suggesting that ROS play an important role in the resolution of inflammation by inducing neutrophil apoptosis. In this experiment, boiled Cu,Zn-SOD had no effect on inhibiting the apoptosis, but boiled Mn-SOD from Bacillus stearothermophilus was more effective in inhibiting the apoptosis than untreated Mn-SOD at the same dose. However, the boiled Mn-SOD showed only 80% of O 2 -inhibitable activity compared with the untreated Mn-SOD. This effect may be attributed to the partial liberation of manganese because MnCl 2 inhibited the apoptosis effectively. Furthermore, Cu,Zn-SOD was effective in delaying apoptosis only when added to the culture within the first 3 h of incubation, suggesting that the isolation of neutrophils from peripheral blood enhances apoptosis of neutrophils.

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Purification and Characterization of the Ca2+ Plus Mg2+-Dependent Endodeoxyribonuclease from Calf Thymus Chromatin1

Cited by 68 publications

References 0 publications

Endogenous endonuclease-induced DNA fragmentation: an early event in cell-mediated cytolysis.

Endogenous endonuclease-induced DNA fragmentation: an early event in cell-mediated cytolysis.

Induction of a Ca2+, Mg2+-dependent endonuclease activity during the early stages of murine erythroleukemic cell differentiation.

Inhibition of Neutrophil Apoptosis by Antioxidants in Culture Medium

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