Characterization of human DNase I family endonucleases and activation of DNase gamma during apoptosis.
We describe here the characterization of the so far identified human DNase I family DNases, DNase I, DNase X, DNase gamma, and DNAS1L2. The DNase I family genes are found to be expressed with different tissue specificities and suggested to play unique physiological roles. All the recombinant DNases are shown to be Ca(2+)/Mg(2+)-dependent endonucleases and catalyze DNA hydrolysis to produce 3'-OH/5'-P ends. High activities for DNase I, DNase X, and DNase gamma are observed under neutral conditions, whereas DNAS1L2 shows its maximum activity at acidic pH. These enzymes have also some other peculiarities: different sensitivities to G-actin, aurintricarboxylic acid, and metal ions are observed. Using a transient expression system in HeLa S3 cells, the possible involvement of the DNases in apoptosis was examined. The ectopic expression of each DNase has no toxic effect on the host cells; however, extensive DNA fragmentation is observed only in DNase gamma-transfected cells after the induction of apoptosis. Furthermore, DNase gamma is revealed to be located at the perinuclear region in living cells, and to translocate into the nucleus during apoptosis. Our results demonstrate that DNase I, DNase X, DNase gamma, and DNAS1L2 have similar but unique endonuclease activities, and that among DNase I family DNases, DNase gamma is capable of producing apoptotic DNA fragmentation in mammalian cells.
Shiokawa
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Department of Biochemistry, Faculty of Pharmaceutical Sciences, Science University of Tokyo, 12 Funagawara-machi, Ichigaya, Shinjuku-ku, Tokyo 162-0826, Japan.
Tanuma
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D000199:Actins / Q000494:pharmacology; D021581:Active Transport, Cell Nucleus / Q000235:genetics; D000328:Adult; D000595:Amino Acid Sequence; D017209:Apoptosis / Q000235:genetics; D002460:Cell Line; D003850:Deoxyribonuclease I / Q000037:antagonists & inhibitors / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D004706:Endodeoxyribonucleases / Q000037:antagonists & inhibitors / Q000096:biosynthesis / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D004789:Enzyme Activation / Q000235:genetics; D004791:Enzyme Inhibitors / Q000494:pharmacology; D006367:HeLa Cells; D006801:Humans; D006868:Hydrolysis; D008969:Molecular Sequence Data; D005810:Multigene Family; D011994:Recombinant Proteins / Q000037:antagonists & inhibitors / Q000096:biosynthesis / Q000737:chemistry; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid; D014162:Transfection; D014960:X Chromosome / Q000201:enzymology
D000199:Actins; D004791:Enzyme Inhibitors; D011994:Recombinant Proteins; D004706:Endodeoxyribonucleases; C105124:deoxyribonuclease gamma; C107519:DNASE1L2 protein, human; D003850:Deoxyribonuclease I