Human urine deoxyribonuclease II (DNase II) isoenzymes: a novel immunoaffinity purification, biochemical multiplicity, genetic heterogeneity and broad distribution among tissues and body fluids

Yasuda, Toshihiro; Nadano, Daita; Awazu, Shuichi; Kishi, Koichiro

doi:10.1016/0167-4838(92)90390-y

Cited by 70 publications

(63 citation statements)

References 25 publications

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“…DNase I is a secretory protein that is highly expressed in the kidney and small intestine, rather than the liver, in rats (49); it also cleaves both the core region and the linker region of chromatin (50). DNase II has been reported to act under acidic conditions (51,52); however, it cleaves chromatin DNA with 3Ј-phosphoryl (P)/5Ј-hydroxyl (OH) ends, which are different from TUNEL-positive cleavage, 3Ј-OH/ 5Ј-P (53). The DNase ␥ proenzyme is normally located in the perinuclear region; apoptotic stimuli process its N-terminal precursor peptide and the mature form is translocated into nuclei, followed by nucleosomal DNA fragmentation (49,54).…”

Section: Discussionmentioning

confidence: 99%

Involvement of Endonuclease G in Nucleosomal DNA Fragmentation under Sustained Endogenous Oxidative Stress

Ishihara

Shimamoto

2006

Journal of Biological Chemistry

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We have previously shown that inhibition of catalase and glutathione peroxidase activities by 3-amino-1,2,4-triazole (ATZ) and mercaptosuccinic acid (MS), respectively, in rat primary hepatocytes caused sustained endogenous oxidative stress and apoptotic cell death without caspase-3 activation. In this study, we investigated the mechanism of this apoptotic cell death in terms of nucleosomal DNA fragmentation. Treatment with ATZ؉MS time-dependently increased the number of deoxynucleotidyl transferase-mediated nick end-labeling (TUNEL)-positive nuclei from 12 h, resulting in clear DNA laddering at 24 h. The deoxyribonuclease (DNase) inhibitor, aurintricarboxylic acid (ATA), completely inhibited nucleosomal DNA fragmentation but the pan-caspase inhibitor, z-VAD-fmk was without effects; furthermore, the cleavage of inhibitor of caspase-activated DNase was not detected, indicating the involvement of DNase(s) other than caspase-activated DNase. Considering that endonuclease G (EndoG) reportedly acts in a caspase-independent manner, we cloned rat EndoG cDNA for the first time. Recombinant EndoG alone digested plasmid DNA and induced nucleosomal DNA fragmentation in isolated hepatocyte nuclei. Recombinant EndoG activity was inhibited by ATA but not by hydrogen peroxide, even at 10 mM. ATZ؉MS stimulation elicited decreases in mitochondrial membrane potential and EndoG translocation from mitochondria to nuclei. By applying RNA interference, the mRNA levels of EndoG were almost completely suppressed and the amount of EndoG protein was decreased to approximately half the level of untreated cells. Under these conditions, decreases in TUNEL-positive nuclei were significantly suppressed. These results indicate that EndoG is responsible, at least in part, for nucleosomal DNA fragmentation under endogenous oxidative stress conditions induced by ATZ؉MS.

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Section: Discussionmentioning

confidence: 99%

Involvement of Endonuclease G in Nucleosomal DNA Fragmentation under Sustained Endogenous Oxidative Stress

Ishihara

Shimamoto

2006

Journal of Biological Chemistry

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“…An antibody against DNase I is commercially available. DNase II antibodies with low titre have been obtained by two laboratories (9,30). Thus, we prepared a polyclonal antibody against DNase II having a high titre, which does not cross-react with DNase I and is able to inhibit DNase II activity in vitro.…”

Section: Discussionmentioning

confidence: 99%

Involvement of DNase II in Nuclear Degeneration during Lens Cell Differentiation

Torriglia¹,

Chaudun²,

F³

et al. 1995

Journal of Biological Chemistry

107

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The characterization of DNase II and DNase I activity was undertaken to discriminate their different roles in physiological nuclear degradation during lens fiber cell differentiation. The activity of both nucleases determined in a new assay allows to discriminate DNase II from DNase I in the same extract. In fibers, both types of nuclease activities are found and appear higher than in epithelial cells. Specific polyclonal antibodies directed against these two nucleases reveal by Western blot analysis the presence of various DNase isoforms. DNase II like-nuclease, present in fibers, is represented by three major bands (60, 23, and 18 kDa), which are not detected, at least for two of them (60 and 23 kDa), in epithelial cells. DNase I like-nuclease pattern in fiber cells shows a single 32-kDa band, while several bands can be detected in epithelial cells. Immunocytochemistry studies show both nucleases present in lens cell sections. DNase II is, as usual, in cytoplasm of epithelial cells, but it appears strikingly concentrated in the nuclei of fibers. DNase I is always concentrated in nuclei of epithelial and fiber cells. DNA degradation observed in agarose gels shows that DNase II-activating medium cleaves the DNA from fiber cells more efficiently than DNase I-activating buffer. In addition, DNase II antibody is able to prevent this degradation. These results suggest a specific involvement of DNase II in nuclear degradation during lens cell differentiation.Apoptosis or programmed cell death occurs in many physiological and pathological situations where selection of cells is required (1-4). In 1980, a landmark study (5) revealed that glucocorticoids induced extensive DNA degradation in rat thymocytes in vitro at the onset of cell death. DNA cleavage occurred in a very specific pattern producing fragments of DNA that were multiples of 180 -200 base pairs. This indicated that the chromatin was cleaved at the linker DNA between nucleosomic cores. The characteristic ladder was first shown by Hewish and Burgoyne (6) To date, three different endonucleases have been involved in DNA fragmentation leading to nucleosomal appearance. Some authors, such as Peitsch et al. (8), claimed that the well characterized pancreatic deoxyribonuclease (DNase I) was constitutively expressed in cells of tissues potentially primed for apoptosis. This 30-kDa nuclease, active at neutral pH, could be responsible for DNA cleavage into oligonucleosomes during cell death. On the other hand, Barry and Eastman (9), studying apoptosis in Chinese hamster ovary cells, were unable to detect a Ca 2ϩ -Mg 2ϩ -dependent endonuclease. Instead, they identified another endonuclease, which was cation-independent, with optimal activity at pH 5. This enzyme was proposed to be DNase II (9, 10). Finally, Hughes and Cidlowski (11) showed a lower molecular weight nuclease, an 18-kDa peptide (termed NUC 18), which was activated by Ca 2ϩ and Mg 2ϩ and related to cyclophilin (12). This peptide appears as a novel enzyme whose activity correlates with apoptosis in thymocy...

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“…DNase I enzymes (EC 3.1.21.1), have a pH optimum at approximately pH 7.0 and requires magnesium ion and many DNaes are purified that are related to DNase 1 family such as DNase1L1, DNase1L2 and DNase1L3 [1,2]. DNase II family (Acid DNase family) (EC 3.1.22.1) are one of the best characterized endonucleases that catalyze DNA hydrolysis in the absence of divalent cations at acidic pH and are found in a wide variety of animal tissues [1,3,4]. Three acid DNases have been identified in mammals so far: DNase 2a, usually referred to as DNase 2, [5], DNase 2b, also known as DLAD [6] and L-DNase II [7].…”

Section: Introductionmentioning

confidence: 99%

A Novel Method For Purifying a DNase From Lysosomal Fraction From Human Small Intestine

Shikara¹,

Al-jaff²

2012

Turk J Bioch

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Aims:The purification of an acid DNase related DNase II family from lysosomes of human small intestine (jejunum). Materials and Methods: Two different methods: a column chromatography series, including phosphocellulose, CM-cellulose and Sephadex G-200 and a novel procedure that use immunoabsorbent technique were used in the purification. Results:The purified DNase is an endonuclease that consisted of one polypeptide chain (monomer) with a molecular mass of 28-32kDa with an optimal pH and temperature of 6.0 and 30°C, respectively. The enzyme prefers native DNA on denatured DNA Discussion: The catalytic properties of the purified enzyme are essentially the same as those of DNase II family such as independency of divalent ions which inhibited its activity at 10mM. The enzyme acts on dsDNA and ssDNA and generates 3'-phosphate and 5'-OH termini which was a characteristic of DNase II. The functions of the intestinal purified DNase from lysosomes were unclear, but lysosomes contained a set of enzymes required for degradation of food, and the enzyme may be necessary for degradation of nucleic acids within food, but small amounts of the enzyme were found in the nuclei and cytoplasm were detected. Several researchers suggested that DNase II family may be active in apoptosis and played a role as a barrier to transfection. The purified was designated SIDNase. Key words: DNase II, small intestine, immunoabsorption, lysosomes, DLAD Conflict of interests: There is no conflict in interest. ÖZETAmaç: İnsan ince bağırsağından (jejunum) asit DNaz II ailesi ile ilintili DNaz izolasyonu. Materyal ve Metod: İzolasyonlar için iki farklı metod uygulanmıştır: Fosfoselüloz CMselüloz ve Sephadex G-200 den oluşan bir kolon serisi ve immünabsorbsiyon tekniği kullanılarak geliştirilen yeni bir prosedür. Sonuçlar: İzole edilen DNaz tek bir polipeptit zincirinden (monomer) oluşan bir endonükleaz olup, moleküler ağırlığı 28-32 kDa, fonksiyonu için optimal ısısı 30°C, pH ise 6.0 dır. Bu enzim natif DNA'yı denatüre DNA'ya tercih etmektedir. Tartışma: İzole edilen enzimin katalitik özellikleri DNaz II ailesine ait enzimler ile temelde aynıdır. Mesela iki değerlikli iyonlardan bağımsız aktivite göstermesi, aktivitesinin 10mM da inhibe olması gibi. Bu enzim dsDNA ve ssDNA üzerinde DNaz II gibi 3'-fosfat ve 5'-OH terminali oluşturur. Bağırsaklardan izole edilen lizozomal DNaz'ın fonksiyonları tam olarak nitelenememiştir ancak lizozomler gıda parçalanması için bir set enzim içermektedir ve gıdada bulunan nükleik asitlerin parçalanması için gerekli olabilirler. Az miktarda enzim çekirdek ve sitoplazmada bulunmuştur. Bazı araştırmacılar DNAz II ailesinin apoptozda aktif olabileceğini ve görevinin transfeksiyonda bariyer oluşturmak olabileceğini öne sürmüşlerdir. İzolat SIDNaz olarak adlandırılmsiyonıştır.

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Human urine deoxyribonuclease II (DNase II) isoenzymes: a novel immunoaffinity purification, biochemical multiplicity, genetic heterogeneity and broad distribution among tissues and body fluids

Cited by 70 publications

References 25 publications

Involvement of Endonuclease G in Nucleosomal DNA Fragmentation under Sustained Endogenous Oxidative Stress

Involvement of Endonuclease G in Nucleosomal DNA Fragmentation under Sustained Endogenous Oxidative Stress

Involvement of DNase II in Nuclear Degeneration during Lens Cell Differentiation

A Novel Method For Purifying a DNase From Lysosomal Fraction From Human Small Intestine

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