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Inhibition of deoxyribonuclease I by actin is to protect cells from premature cell death

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Abstract

Deoxyribonuclease I (Dnase1) is the major extracellular endonuclease. It is secreted by digestive glands into the alimentary tract and into the plasma, lacrimal fluid and urine by hepatocytes, lacrimal glands and renal proximal tubular cells, respectively. In many species the activity of Dnase1 is inhibited by monomeric actin. However, the biological significance of this high affinity interaction is unknown. We generated a Dnase1 mutant with extremely reduced actin binding capacity. EGFP-constructs of wild-type and mutant Dnase1 were transfected into MCF-7 breast cancer cells and apoptosis or necrosis was induced by staurosporine or oxidative stress. During apoptosis faster chromatin fragmentation occurred in cells transfected with mutant Dnase1. When wt (wild-type)- or mutated Dnase1 were added to cells after induction of necrosis, faster chromatin degradation occurred in the presence of mutant Dnase1. Inclusion of actin under these conditions inhibited chromatin degradation by wt- but not by mutated Dnase1. Thus, inhibition of Dnase1 by actin may serve as a self-protection mechanism against premature DNA degradation during cell damage.

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References

  1. Sachs F (1905) Über die Nuclease. Hoppe Sylers Z Phys Chem 46:337–353

    CAS  Google Scholar 

  2. Kunitz M (1950) Crystalline deoxyribonuclease. J Gen Physiol 33:349–362

    Article  PubMed  CAS  Google Scholar 

  3. Kreuder V, Dieckhoff J, Sittig M, Mannherz HG (1984) Isolation, characterisation and crystallization of deoxyribonuclease I from bovine and rat parotid gland and its interaction with rabbit skeletal muscle actin. Eur J Biochem 139:389–400

    Article  PubMed  CAS  Google Scholar 

  4. Mannherz HG, Peitsch MC, Zanotti S, Paddenberg R, Polzar B (1995) A new function for an old enzyme: the role of DNase I in apoptosis. Curr Top Microbiol Immunol 198:161–174

    PubMed  CAS  Google Scholar 

  5. Takeshita H, Yasuda T, Nakajima T, Hosomi O, Nakashima Y, Kishi K (1997) Mouse deoxyribonuclease I (DNase I): biochemical and immunological characterization, cDNA structure and tissue distribution. Biochem Mol Biol Int 42:65–75

    PubMed  CAS  Google Scholar 

  6. Napirei M, Gültekin A, Kloeckl T, Möröy T, Frostegard J, Mannherz HG (2006) Systemic lupus-erythematosus: deoxyribonuclease 1 in necrotic chromatin disposal. Int J Biochem Cell Biol 38:297–306

    Article  PubMed  CAS  Google Scholar 

  7. Lindberg MU (1966) Crystallization from calf spleen of two inhibitors of deoxyribonuclease I. J Biol Chem 241:1246–1249

    PubMed  CAS  Google Scholar 

  8. Lazarides E, Lindberg U (1974) Actin is the naturally occurring inhibitor of deoxyribonuclease I. Proc Natl Acad Sci USA 71:4742–4746

    Article  PubMed  CAS  Google Scholar 

  9. Mannherz HG, Leigh JB, Leberman R, Pfrang H (1975) A specific 1:1 G-actin:DNAase I complex formed by the action of DNAase I on F-actin. FEBS Lett 60:34–38

    Article  PubMed  CAS  Google Scholar 

  10. Hitchcock SE, Carlsson L, Lindberg U (1976) Depolymerization of F-actin by deoxyribonuclease I. Cell 7:531–542

    Article  PubMed  CAS  Google Scholar 

  11. Mannherz HG, Goody RS, Konrad M, Nowak E (1980) The interaction of bovine pancreatic deoxyribonuclease I and skeletal muscle actin. Eur J Biochem 104:367–379

    Article  PubMed  CAS  Google Scholar 

  12. Mannherz HG (1992) Crystallization of actin in complex with actin-binding proteins. J Biol Chem 267:11661–11664

    PubMed  CAS  Google Scholar 

  13. Kothakota S, Azuma T, Reinhard C et al (1997) Caspase-3-generated fragment of gelsolin: effector of morphological change in apoptosis. Science 278:294–298

    Article  PubMed  CAS  Google Scholar 

  14. Chhabra D, Nosworthy NJ, dos Remedios CG (2005) The N-terminal fragment of gelsolin inhibits the interaction of DNase I with isolated actin, but not with the cofilin-actin complex. Proteomics 5:3131–3136

    Article  PubMed  CAS  Google Scholar 

  15. Polzar B, Nowak E, Goody RS, Mannherz HG (1989) The complex of actin and deoxyribonuclease I as a model system to study the interactions of nucleotides, cations and cytochalasin D with monomeric actin. Eur J Biochem 182:267–275

    Article  PubMed  CAS  Google Scholar 

  16. Blikstad I, Markey F, Carlsson L, Persson T, Lindberg U (1978) Selective assay of monomeric and filamentous actin in cell extracts, using inhibition of deoxyribonuclease I. Cell 15:935–943

    Article  PubMed  CAS  Google Scholar 

  17. Lind SE, Smith DB, Janmey PA, Stossel TP (1988) Depression of gelsolin levels and detection of gelsolin-actin complexes in plasma of patients with acute lung injury. Am Rev Respir Dis 138:429–434

    PubMed  CAS  Google Scholar 

  18. Kabsch W, Mannherz HG, Suck D (1985) Three-dimensional structure of the complex of actin and DNase I at 4.5 A resolution. EMBO J 4:2113–2118

    PubMed  CAS  Google Scholar 

  19. Hosomi O, Yasuda T, Takeshita H et al (2000) Molecular cloning of cDNA encoding Xenopus laevis deoxyribonuclease I. DNA Seq 11:247–255

    PubMed  CAS  Google Scholar 

  20. Mori S, Yasuda T, Takeshita H et al (2001) Molecular, biochemical and immunological analyses of porcine pancreatic DNase I. Biochim Biophys Acta 1547:275–287

    PubMed  CAS  Google Scholar 

  21. Nakashima Y, Yasuda T, Takeshita H et al (1999) Molecular, biochemical and immunological studies of hen pancreatic deoxyribonuclease I. Int J Biochem Cell Biol 31:1315–1326

    Article  PubMed  CAS  Google Scholar 

  22. Hitchcock SE (1980) Actin deoxyroboncuclease I interaction. Depolymerization and nucleotide exchange. J Biol Chem 255:5668–5673

    PubMed  CAS  Google Scholar 

  23. Ulmer JS, Herzka A, Toy KJ et al (1996) Engineering actin-resistant human DNase I for treatment of cystic fibrosis. Proc Natl Acad Sci USA 93:8225–8229

    Article  PubMed  CAS  Google Scholar 

  24. Fujihara J, Hieda Y, Xue Y et al (2006) Actin-inhibition and folding of vertebrate deoxyribonuclease I are affected by mutations at residues 67 and 114. Comp Biochem Physiol B Biochem Mol Biol 143:70–75

    Article  PubMed  CAS  Google Scholar 

  25. Pan CQ, Ulmer JS, Herzka A, Lazarus RA (1998) Mutational analysis of human DNase I at the DNA binding interface: implications for DNA recognition, catalysis, and metal ion dependence. Protein Sci 7:628–636

    Article  PubMed  CAS  Google Scholar 

  26. Lacks SA (1981) Deoxyribonuclease I in mammalian tissues. Specificity of inhibition by actin. J Biol Chem 256:2644–2648

    PubMed  CAS  Google Scholar 

  27. Shiokawa D, Tanuma S (2001) Characterization of human DNase I family endonucleases and activation of DNase gamma during apoptosis. Biochemistry 40:143–152

    Article  PubMed  CAS  Google Scholar 

  28. Napirei M, Karsunky H, Zevnik B, Stephan H, Mannherz HG, Möröy T (2000) Features of systemic lupus erythematosus in Dnase1-deficient mice. Nat Genet 25:177–181

    Article  PubMed  CAS  Google Scholar 

  29. Chitrabamrung S, Rubin RL, Tan EM (1981) Serum deoxyribonuclease I and clinical activity in systemic lupus erythematosus. Rheumatol Int 1:55–60

    Article  PubMed  CAS  Google Scholar 

  30. Macanovic M, Lachmann PJ (1997) Measurement of deoxyribonuclease I (DNase) in the serum and urine of systemic lupus erythematosus (SLE)-prone NZB/NZW mice by a new radial enzyme diffusion assay. Clin Exp Immunol 108:220–226

    Article  PubMed  CAS  Google Scholar 

  31. Nagata S (2005) DNA degradation in development and programmed cell death. Annu Rev Immunol 23:853–875

    Article  PubMed  CAS  Google Scholar 

  32. Napirei M, Wulf S, Mannherz HG (2004) Chromatin breakdown during necrosis by serum Dnase 1 and the plasminogen system. Arthritis Rheum 50:1873–1883

    Article  PubMed  CAS  Google Scholar 

  33. Ho SN, Hunt HD, Horton RM, Pullen JK, Pease LR (1989) Site-directed mutagenesis by overlap extension using the polymerase chain reaction. Gene 77:51–59

    Article  PubMed  CAS  Google Scholar 

  34. Ballweber E, Galla M, Aktories K, Yeoh S, Weeds AG, Mannherz HG (2001) Interaction of ADP-ribosylated actin with actin binding proteins. FEBS Lett 508:131–135

    Article  PubMed  CAS  Google Scholar 

  35. Mclaughlin PJ, Gooch JT, Mannherz HG, Weeds AG (1993) Structure of gelsolin segment-1-actin complex and the mechanism of filament severing. Nature 364:685–692

    Article  PubMed  CAS  Google Scholar 

  36. Napirei M, Ricken A, Eulitz D, Knoop H, Mannherz HG (2004) Expression pattern of the deoxyribonuclease 1 gene: lessons from the Dnase 1 knockout mouse. Biochem J 380:929–937

    Article  PubMed  CAS  Google Scholar 

  37. Nadano D, Yasuda T, Kishi K (1993) Measurement of deoxyribonuclease I activity in human tissues and body fluids by a single radial enzyme-diffusion method. Clin Chem 39:448–452

    PubMed  CAS  Google Scholar 

  38. Kabsch W, Mannherz HG, Suck D, Pai EF, Holmes KC (1990) Atomic structure of the actin:DNase I complex. Nature 347:37–44

    Article  PubMed  CAS  Google Scholar 

  39. Weston SA, Lahm A, Suck D (1992) X-ray structure of the DNase I-d(GGTATACC)2 complex at 2.3 A resolution. J Mol Biol 226:1237–1256

    Article  PubMed  CAS  Google Scholar 

  40. Ballweber E, Hannappel E, Huff T, Mannherz HG (1997) Mapping the binding site of thymosin beta4 on actin by competition with G-actin binding proteins indicates negative co-operativity between binding sites located on opposite subdomains of actin. Biochem J 327:787–793

    PubMed  CAS  Google Scholar 

  41. Oberhammer F, Wilson JW, Dive C et al (1993) Apoptotic death in epithelial cells: cleavage of DNA to 300 and/or 50 kb fragments prior to or in the absence of internucleosomal fragmentation. EMBO J 12:3679–3684

    PubMed  CAS  Google Scholar 

  42. Otto AM, Paddenberg R, Schubert S, Mannherz HG (1996) Cell-cycle arrest, micronucleus formation, and cell death in growth inhibition of MCF-7 breast cancer cells by tamoxifen and cisplatin. J Cancer Res Clin Oncol 122:603–612

    Article  PubMed  CAS  Google Scholar 

  43. Kerr JF, Wyllie AH, Currie AR (1972) Apoptosis: a basic biological phenomenon with wide-ranging implications in tissue kinetics. Br J Cancer 26:239–257

    PubMed  CAS  Google Scholar 

  44. Walport MJ (2000) Lupus, DNase and defective disposal of cellular debris. Nat Genet 25:135–136

    Article  PubMed  CAS  Google Scholar 

  45. Basnakian AG, Apostolov EO, Yin X, Napirei M, Mannherz HG, Shah SV (2005) Cisplatin nephrotoxicity is mediated by deoxyribonuclease I. J Am Soc Nephrol 16:697–702

    Article  PubMed  CAS  Google Scholar 

  46. Napirei M, Basnakian AG, Apostolov EO, Mannherz HG (2006) Deoxyribonuclease 1 aggravates acetaminophen-induced liver necrosis in male CD-1 mice. Hepatology 43:297–305

    Article  PubMed  CAS  Google Scholar 

  47. Peitsch MC, Polzar B, Stephan H et al (1993) Characterization of the endogenous deoxyribonuclease involved in nuclear-dna degradation during apoptosis (programmed cell-death). EMBO J 12:371–377

    PubMed  CAS  Google Scholar 

  48. Superti F, Ammendolia MG, Tinari A et al (1996) Induction of apoptosis in HT-29 cells infected with SA-11 rotavirus. J Med Virol 50:325–334

    Article  PubMed  CAS  Google Scholar 

  49. Mannherz HG, Ballweber E, Galla M et al (2007) Mapping the ADF/Cofilin binding site on monomeric actin by competitive cross-linking and peptide array: evidence for a second binding site on monomeric actin. J Mol Biol 366:745–755

    Article  PubMed  CAS  Google Scholar 

  50. Nosworthy NJ, Kekic M, dos Remedios CG (2001) The affinity of chick cofilin for actin increases when actin is complexed with DNase I: formation of a cofilin-actin-DNase I ternary complex. Proteomics 1:1513–1518

    Article  PubMed  CAS  Google Scholar 

  51. Davies KE, Nowak KJ (2006) Molecular mechanisms of muscular dystrophies: old and new players. Nat Rev Mol Cell Biol 7:762–773

    Article  PubMed  CAS  Google Scholar 

  52. Manderson AP, Carlucci F, Lachmann PJ et al (2006) The in vivo expression of actin/salt-resistant hyperactive DNase I inhibits the development of anti-ssDNA and anti-histone autoantibodies in a murine model of systemic lupus erythematosus. Arthritis Res Ther 8:R68

    Google Scholar 

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Acknowledgement

We thank Mrs. E.-M. Konieczny, K. Klar, R. Ritenberg, and S. Wulf for technical assistance.

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  1. Department of Anatomy and Embryology, Ruhr-University Bochum, Universitätsstraße 150, 44780, Bochum, Germany

    Dirk Eulitz & Hans Georg Mannherz

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  1. Dirk Eulitz
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  2. Hans Georg Mannherz
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Correspondence to Dirk Eulitz.

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Eulitz, D., Mannherz, H.G. Inhibition of deoxyribonuclease I by actin is to protect cells from premature cell death. Apoptosis 12, 1511–1521 (2007). https://doi.org/10.1007/s10495-007-0078-4

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