Action of apoptotic endonuclease DNase γ on naked DNA and chromatin substrates

Evaluation of all non‐synonymous single nucleotide polymorphisms (SNPs) in the genes encoding human deoxyribonuclease I and I‐like 3 as a functional SNP potentially implicated in autoimmunity

“…Endogenous deoxyribonuclease I (DNase I, EC3.1.21.1) and DNase I-like 3 (DNase 1L3) have been noted as candidate endonucleases involved in the breakdown of chromatin during apoptosis and/or necrosis [1][2][3]. DNase 1L3 is a member of the DNase I family, which also includes DNase I-like 1/DNase X/DNase Xib and DNase I-like 2, whose nucleotide and amino acid sequences resemble those of the originally characterized DNase I [4,5].…”

Possible contribution of DNase γ to immunoglobulin V gene diversification

“…On the other hand, the generation of GC was independent of DNase ␥ expression. Given the staggered DSBs are raised by single-strand breaks occurring in proximity on both strands, DNase ␥, preferably cutting single-strand DNA in a physiological ionic strength in vitro [23], may be directly involved in the generation of such DSBs. These results together suggest that DNase ␥-mediated staggered DSBs in the V region gene is at least partly involved in the machinery of the non-templated mutations of this region.…”

“…DNase ␥ is highly expressed in lymphoid organs and selectively in germinal center B cells, and its expression markedly increases in B cells upon CD40 stimulation in parallel with AID expression [19]. DNase ␥ is responsible for nucleosomal DNA fragmentation in some instances of apoptosis [21,22], although it preferably cuts a single-strand portion of naked DNA in vitro at physiological ionic strength [23]. Despite its well-defined enzymatic nature, physiological role of DNase ␥ in vivo remains unknown.…”

DNase γ-dependent and -independent apoptotic DNA fragmentations in Ramos Burkitt's lymphoma cell line

“…DNase γ is located in the nuclear envelope in living cells and translocates into the nucleus in dying cells, where it induces DNA fragmentation (17). Although some of the catalytic properties of DNase γ have been reported (17,10), the role of Isolation of nuclei and induction of DNA fragmentation. Cells were suspended and lysed in a hypotonic buffer (10 mM Hepes, 10 mM KCl, 3 mM MgCl 2 , 0.5 mM spermidine, and 0.1% NP40) for 5 min on ice and nuclei were harvested by centrifugation.…”

“…Then, the isolated nuclei (5 × 10 5 ) were suspended in 60 μL of a buffer containing 25 mM Tris-HCl (pH 7.4), 150 mM KCl, and 5 mM MgCl 2 , with or without 5 mM CaCl 2 . The reaction was performed at 37°C for 1 h with or without 1 ng of the recombinant DNase γ (10,19). DNA was purified from the nuclei with Wizard DNA purification resin (Promega, Madison, WI, USA) as described previously (21) and DNA fragmentation was analyzed by electrophoresis in a 1.5% agarose gel containing ethidium bromide (0.1 μg/mL).…”