Adenosine, deoxyadenosine, and deoxyguanosine induce DNA cleavage in mouse thymocytes.
When thymocytes were cultured with adenosine, deoxyadenosine, or deoxyguanosine at 1 mM for 24 h, DNA cleavage at internucleosomal sites with multiples of approximately 180 bp was induced, followed by lactate dehydrogenase release into the medium. In the presence of coformycin, an adenosine deaminase inhibitor, or formycin B, a purine nucleoside phosphorylase inhibitor, DNA cleavage was induced by these nucleosides at concentrations of less than 50 microM. Other purine and pyrimidine ribo- and deoxyribonucleosides did not induce DNA cleavage or LDH release. Because thymocyte nuclei contain a Ca2+,Mg2+-dependent endonuclease, which preferentially cuts DNA in its linker regions, DNA fragmentation induced by the three purine nucleosides was suggested to occur through increased activity of the endonuclease. The DNA cleavage induced by the nucleosides required protein phosphorylation and synthesis, inasmuch as it was inhibited by an inhibitor of protein kinases, H-7, and by an inhibitor of protein synthesis, cycloheximide. The inhibition of DNA cleavage was accompanied by a reduction in lactate dehydrogenase release, suggesting a causal relationship between DNA cleavage and cell death. The DNA cleavage and subsequent cell lysis might be related to the selective thymocyte deletion observed in patients with adenosine deaminase or purine nucleoside phosphorylase deficiency.
Kizaki
H
H
Department of Biochemistry, School of Medicine, Keio University, Tokyo, Japan.
Shimada
H
H
Ohsaka
F
F
Sakurada
T
T
D019307:1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine; D000241:Adenosine / Q000633:toxicity; D000818:Animals; D002118:Calcium / Q000494:pharmacology; D003513:Cycloheximide / Q000494:pharmacology; D004247:DNA / Q000302:isolation & purification; D004249:DNA Damage / Q000187:drug effects; D003839:Deoxyadenosines / Q000633:toxicity; D003849:Deoxyguanosine / Q000633:toxicity; D007546:Isoquinolines / Q000494:pharmacology; D007700:Kinetics; D007770:L-Lactate Dehydrogenase / Q000378:metabolism; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D010879:Piperazines / Q000494:pharmacology; D011064:Poly Adenosine Diphosphate Ribose / Q000037:antagonists & inhibitors; D011684:Purine Nucleosides / Q000633:toxicity*; D013601:T-Lymphocytes / Q000187:drug effects* / Q000378:metabolism / Q000502:physiology; D015032:Zinc / Q000494:pharmacology
D003839:Deoxyadenosines; D007546:Isoquinolines; D010879:Piperazines; D011684:Purine Nucleosides; D011064:Poly Adenosine Diphosphate Ribose; D019307:1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine; D004247:DNA; D003513:Cycloheximide; D007770:L-Lactate Dehydrogenase; D003849:Deoxyguanosine; D015032:Zinc; D000241:Adenosine; D002118:Calcium