Abstract
The tissue distribution of deoxyribonuclease 1 (DNASE1, DNase I), a Ca2+ and Mg2+/Mn2+-dependent secretory endonuclease, has previously been investigated. However, most of these studies did not account for the existence of different members of the DNASE1 gene family, did not differentiate between endogenous DNASE1 protein synthesis and its extracellular occurrence or were not performed with methods allowing both a sensitive and a specific detection. Now we re-examined the DNASE1 gene expression pattern by taking advantage of the Dnase1 knockout mouse model. Direct comparison of samples derived from wild-type (Dnase1+/+) and knockout (Dnase1-/-) mice allowed an unambiguous detection of Dnase1 gene expression at the mRNA and protein level. For the detection of Dnase1 activity, we developed a highly sensitive nuclease zymogram method. We observed high Dnase1 gene expression in the parotid and submandibular gland as well as in the kidney and duodenum, intermediate expression in the ileum, mesenterial lymph nodes, liver, ventral prostate, epididymis, ovary and stomach, and low expression in the sublingual, preputial, coagulation and pituitary gland. We report for the first time the lachrymal and thyroid glands, the urinary bladder and the eye to be Dnase1-expressing organs as well. Since Dnase1 knockout mice with the 129xC57Bl/6 mixed genetic background have indicated the protection against an anti-DNA autoimmune response as a new physiological function of Dnase1, knowledge of the physiological sites of its synthesis might prove helpful to find new therapeutic strategies.
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Selected References
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- Abe A., Liao T. H. The immunological and structural comparisons of deoxyribonucleases I. Glycosylation differences between bovine pancreatic and parotid deoxyribonucleases. J Biol Chem. 1983 Sep 10;258(17):10283–10288. [PubMed] [Google Scholar]
- Baron W. F., Pan C. Q., Spencer S. A., Ryan A. M., Lazarus R. A., Baker K. P. Cloning and characterization of an actin-resistant DNase I-like endonuclease secreted by macrophages. Gene. 1998 Jul 30;215(2):291–301. doi: 10.1016/s0378-1119(98)00281-9. [DOI] [PubMed] [Google Scholar]
- Boone D. L., Tsang B. K. Identification and localization of deoxyribonuclease I in the rat ovary. Biol Reprod. 1997 Oct;57(4):813–821. doi: 10.1095/biolreprod57.4.813. [DOI] [PubMed] [Google Scholar]
- Bradford M. M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976 May 7;72:248–254. doi: 10.1006/abio.1976.9999. [DOI] [PubMed] [Google Scholar]
- Cacia J., Quan C. P., Pai R., Frenz J. Human DNase I contains mannose 6-phosphate and binds the cation-independent mannose 6-phosphate receptor. Biochemistry. 1998 Oct 27;37(43):15154–15161. doi: 10.1021/bi981465t. [DOI] [PubMed] [Google Scholar]
- Coy J. F., Velhagen I., Himmele R., Delius H., Poustka A., Zentgraf H. Isolation, differential splicing and protein expression of a DNase on the human X chromosome. Cell Death Differ. 1996 Apr;3(2):199–206. [PubMed] [Google Scholar]
- Devillers-Thiery A., Kindt T., Scheele G., Blobel G. Homology in amino-terminal sequence of precursors to pancreatic secretory proteins. Proc Natl Acad Sci U S A. 1975 Dec;72(12):5016–5020. doi: 10.1073/pnas.72.12.5016. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Funakoshi A., Tsubota Y., Wakasugi H., Ibayashi H., Takagi Y. Purification and properties of human pancreatic deoxyribonuclease I. J Biochem. 1977 Dec;82(6):1771–1777. doi: 10.1093/oxfordjournals.jbchem.a131875. [DOI] [PubMed] [Google Scholar]
- Jacob M., Napirei M., Ricken A., Dixkens C., Mannherz H. G. Histopathology of lupus-like nephritis in Dnase1-deficient mice in comparison to NZB/W F1 mice. Lupus. 2002;11(8):514–527. doi: 10.1191/0961203302lu242oa. [DOI] [PubMed] [Google Scholar]
- Koizumi T. Tissue distribution of deoxyribonuclease I (DNase I) activity level in mice and its sexual dimorphism. Exp Anim. 1995 Jul;44(3):181–185. doi: 10.1538/expanim.44.181. [DOI] [PubMed] [Google Scholar]
- Kraehenbuhl J. P., Racine L., Jamieson J. D. Immunocytochemical localization of secretory proteins in bovine pancreatic exocrine cells. J Cell Biol. 1977 Feb;72(2):406–423. doi: 10.1083/jcb.72.2.406. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Kreuder V., Dieckhoff J., Sittig M., Mannherz H. G. Isolation, characterisation and crystallization of deoxyribonuclease I from bovine and rat parotid gland and its interaction with rabbit skeletal muscle actin. Eur J Biochem. 1984 Mar 1;139(2):389–400. doi: 10.1111/j.1432-1033.1984.tb08018.x. [DOI] [PubMed] [Google Scholar]
- Lachmann P. J. Lupus and desoxyribonuclease. Lupus. 2003;12(3):202–206. doi: 10.1191/0961203303lu357xx. [DOI] [PubMed] [Google Scholar]
- Lacks S. A. Deoxyribonuclease I in mammalian tissues. Specificity of inhibition by actin. J Biol Chem. 1981 Mar 25;256(6):2644–2648. [PubMed] [Google Scholar]
- Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
- Liu Q. Y., Pandey S., Singh R. K., Lin W., Ribecco M., Borowy-Borowski H., Smith B., LeBlanc J., Walker P. R., Sikorska M. DNaseY: a rat DNaseI-like gene coding for a constitutively expressed chromatin-bound endonuclease. Biochemistry. 1998 Jul 14;37(28):10134–10143. doi: 10.1021/bi9800597. [DOI] [PubMed] [Google Scholar]
- Los M., Neubüser D., Coy J. F., Mozoluk M., Poustka A., Schulze-Osthoff K. Functional characterization of DNase X, a novel endonuclease expressed in muscle cells. Biochemistry. 2000 Jun 27;39(25):7365–7373. doi: 10.1021/bi000158w. [DOI] [PubMed] [Google Scholar]
- Mannherz H. G., Kreuder V., Koch J., Dieckhoff J., Drenckhahn D. The inhibition of bovine and rat parotid deoxyribonuclease I by skeletal muscle actin. A biochemical and immunocytochemical study. Biochem J. 1982 Nov 1;207(2):305–313. doi: 10.1042/bj2070305. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Mannherz H. G., Peitsch M. C., Zanotti S., Paddenberg R., Polzar B. A new function for an old enzyme: the role of DNase I in apoptosis. Curr Top Microbiol Immunol. 1995;198:161–174. doi: 10.1007/978-3-642-79414-8_10. [DOI] [PubMed] [Google Scholar]
- Nadano D., Yasuda T., Kishi K. Measurement of deoxyribonuclease I activity in human tissues and body fluids by a single radial enzyme-diffusion method. Clin Chem. 1993 Mar;39(3):448–452. [PubMed] [Google Scholar]
- Napirei M., Karsunky H., Zevnik B., Stephan H., Mannherz H. G., Möröy T. Features of systemic lupus erythematosus in Dnase1-deficient mice. Nat Genet. 2000 Jun;25(2):177–181. doi: 10.1038/76032. [DOI] [PubMed] [Google Scholar]
- Nishikawa A., Mizuno S. The efficiency of N-linked glycosylation of bovine DNase I depends on the Asn-Xaa-Ser/Thr sequence and the tissue of origin. Biochem J. 2001 Apr 1;355(Pt 1):245–248. doi: 10.1042/0264-6021:3550245. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Parrish J. E., Ciccodicola A., Wehhert M., Cox G. F., Chen E., Nelson D. L. A muscle-specific DNase I-like gene in human Xq28. Hum Mol Genet. 1995 Sep;4(9):1557–1564. doi: 10.1093/hmg/4.9.1557. [DOI] [PubMed] [Google Scholar]
- Pergolizzi R., Appierto V., Bosetti A., DeBellis G. L., Rovida E., Biunno I. Cloning of a gene encoding a DNase I-like endonuclease in the human Xq28 region. Gene. 1996 Feb 12;168(2):267–270. doi: 10.1016/0378-1119(95)00741-5. [DOI] [PubMed] [Google Scholar]
- Polzar B., Zanotti S., Stephan H., Rauch F., Peitsch M. C., Irmler M., Tschopp J., Mannherz H. G. Distribution of deoxyribonuclease I in rat tissues and its correlation to cellular turnover and apoptosis (programmed cell death). Eur J Cell Biol. 1994 Jun;64(1):200–210. [PubMed] [Google Scholar]
- Price P. A., Liu T. Y., Stein W. H., Moore S. Properties of chromatographically purified bovine pancreatic deoxyribonuclease. J Biol Chem. 1969 Feb 10;244(3):917–923. [PubMed] [Google Scholar]
- Rauch F., Polzar B., Stephan H., Zanotti S., Paddenberg R., Mannherz H. G. Androgen ablation leads to an upregulation and intranuclear accumulation of deoxyribonuclease I in rat prostate epithelial cells paralleling their apoptotic elimination. J Cell Biol. 1997 May 19;137(4):909–923. doi: 10.1083/jcb.137.4.909. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Rodriguez A. M., Rodin D., Nomura H., Morton C. C., Weremowicz S., Schneider M. C. Identification, localization, and expression of two novel human genes similar to deoxyribonuclease I. Genomics. 1997 Jun 15;42(3):507–513. doi: 10.1006/geno.1997.4748. [DOI] [PubMed] [Google Scholar]
- Rudolph F. B. The biochemistry and physiology of nucleotides. J Nutr. 1994 Jan;124(1 Suppl):124S–127S. doi: 10.1093/jn/124.suppl_1.124S. [DOI] [PubMed] [Google Scholar]
- Shimada O., Ishikawa H., Tosaka-Shimada H., Yasuda T., Kishi K., Suzuki S. Detection of deoxyribonuclease I along the secretory pathway in Paneth cells of human small intestine. J Histochem Cytochem. 1998 Jul;46(7):833–840. doi: 10.1177/002215549804600706. [DOI] [PubMed] [Google Scholar]
- Shimada O., Suzuki S., Tosaka-Shimada H., Ishikawa H. Detection of deoxyribonuclease I in a hormone-secretory pathway of pituitary cells in humans and rats. Cell Struct Funct. 1998 Apr;23(2):49–56. doi: 10.1247/csf.23.49. [DOI] [PubMed] [Google Scholar]
- Shiokawa D., Tanuma S. Characterization of human DNase I family endonucleases and activation of DNase gamma during apoptosis. Biochemistry. 2001 Jan 9;40(1):143–152. doi: 10.1021/bi001041a. [DOI] [PubMed] [Google Scholar]
- Shiokawa D., Tanuma S. Molecular cloning and expression of a cDNA encoding an apoptotic endonuclease DNase gamma. Biochem J. 1998 Jun 15;332(Pt 3):713–720. doi: 10.1042/bj3320713. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Stephan H., Polzar B., Rauch F., Zanotti S., Ulke C., Mannherz H. G. Distribution of deoxyribonuclease I (DNase I) and p53 in rat testis and their correlation with apoptosis. Histochem Cell Biol. 1996 Oct;106(4):383–393. doi: 10.1007/BF02473297. [DOI] [PubMed] [Google Scholar]
- Takeshita H., Mogi K., Yasuda T., Nakajima T., Nakashima Y., Mori S., Hoshino T., Kishi K. Mammalian deoxyribonucleases I are classified into three types: pancreas, parotid, and pancreas-parotid (mixed), based on differences in their tissue concentrations. Biochem Biophys Res Commun. 2000 Mar 16;269(2):481–484. doi: 10.1006/bbrc.2000.2300. [DOI] [PubMed] [Google Scholar]
- Takeshita H., Yasuda T., Nakajima T., Hosomi O., Nakashima Y., Kishi K. Mouse deoxyribonuclease I (DNase I): biochemical and immunological characterization, cDNA structure and tissue distribution. Biochem Mol Biol Int. 1997 Jun;42(1):65–75. doi: 10.1080/15216549700202441. [DOI] [PubMed] [Google Scholar]
- Tanuma S., Shiokawa D. Multiple forms of nuclear deoxyribonuclease in rat thymocytes. Biochem Biophys Res Commun. 1994 Sep 15;203(2):789–797. doi: 10.1006/bbrc.1994.2252. [DOI] [PubMed] [Google Scholar]
- Tsutsumi S., Kaneko Y., Asao T., Kuwano H., Kudo S., Takeshita H., Yasuda T., Kishi K. DNase I is present in the chief cells of human and rat stomachs. Histochem J. 2001 Sep-Oct;33(9-10):531–535. doi: 10.1023/a:1014999624430. [DOI] [PubMed] [Google Scholar]
- Yasuda T., Takeshita H., Nakajima T., Hosomi O., Nakashima Y., Kishi K. Rabbit DNase I: purification from urine, immunological and proteochemical characterization, nucleotide sequence, expression in tissues, relationships with other mammalian DNases I and phylogenetic analysis. Biochem J. 1997 Jul 15;325(Pt 2):465–473. doi: 10.1042/bj3250465. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Zanotti S., Polzar B., Stephan H., Doll U., Niessing J., Mannherz H. G. Localization of deoxyribonuclease I gene transcripts and protein in rat tissues and its correlation with apoptotic cell elimination. Histochem Cell Biol. 1995 May;103(5):369–377. doi: 10.1007/BF01457812. [DOI] [PubMed] [Google Scholar]
- Zeng Z., Parmelee D., Hyaw H., Coleman T. A., Su K., Zhang J., Gentz R., Ruben S., Rosen C., Li Y. Cloning and characterization of a novel human DNase. Biochem Biophys Res Commun. 1997 Feb 13;231(2):499–504. doi: 10.1006/bbrc.1996.5923. [DOI] [PubMed] [Google Scholar]