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. 2001 Apr 15;355(Pt 2):271–278. doi: 10.1042/0264-6021:3550271

Transgenic mice with inactive alleles for procollagen N-proteinase (ADAMTS-2) develop fragile skin and male sterility.

S W Li 1, M Arita 1, A Fertala 1, Y Bao 1, G C Kopen 1, T K Långsjö 1, M M Hyttinen 1, H J Helminen 1, D J Prockop 1
PMCID: PMC1221736  PMID: 11284712

Abstract

Transgenic mice were prepared with inactive alleles for procollagen N-proteinase (ADAMTS-2; where ADAMTS stands for a disintegrin and metalloproteinase with thrombospondin repeats). Homozygous mice were grossly normal at birth, but after 1-2 months they developed thin skin that tore after gentle handling. Although the gene was inactivated, a large fraction of the N-propeptides of type I procollagen in skin and the N-propeptides of type II procollagen in cartilage were cleaved. Therefore the results suggested the tissues contained one or more additional enzymes that slowly process the proteins. Electron microscopy did not reveal any defects in the morphology of collagen fibrils in newborn mice. However, in two-month-old mice, the collagen fibrils in skin were seen as bizarre curls in cross-section and the mean diameters of the fibrils were approx. half of the controls. Although a portion of the N-propeptides of type II procollagen in cartilage were not cleaved, no defects in the morphology of the fibrils were seen by electron microscopy or by polarized-light microscopy. Female homozygous mice were fertile, but male mice were sterile with a marked decrease in testicular sperm. Therefore the results indicated that ADAMTS-2 plays an essential role in the maturation of spermatogonia.

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Selected References

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  1. Abbaszade I., Liu R. Q., Yang F., Rosenfeld S. A., Ross O. H., Link J. R., Ellis D. M., Tortorella M. D., Pratta M. A., Hollis J. M. Cloning and characterization of ADAMTS11, an aggrecanase from the ADAMTS family. J Biol Chem. 1999 Aug 13;274(33):23443–23450. doi: 10.1074/jbc.274.33.23443. [DOI] [PubMed] [Google Scholar]
  2. Adham I. M., Kim Y., Shamsadin R., Heinlein U. A., Von Beust G., Mattei M. G., Engel W. Molecular cloning, chromosomal localization, and expression analysis of CYRN1 and CYRN2, two human genes coding for cyritestin, a sperm protein involved in gamete interaction. DNA Cell Biol. 1998 Feb;17(2):161–168. doi: 10.1089/dna.1998.17.161. [DOI] [PubMed] [Google Scholar]
  3. Berger J., Tanzawa K., Prockop D. J. Sequential cleavage of type I procollagen by procollagen N-proteinase. An intermediate containing an uncleaved pro alpha 1(I) chain. Biochemistry. 1985 Jan 29;24(3):600–605. doi: 10.1021/bi00324a009. [DOI] [PubMed] [Google Scholar]
  4. Bronson R. A., Fusi F. M., Calzi F., Doldi N., Ferrari A. Evidence that a functional fertilin-like ADAM plays a role in human sperm-oolemmal interactions. Mol Hum Reprod. 1999 May;5(5):433–440. doi: 10.1093/molehr/5.5.433. [DOI] [PubMed] [Google Scholar]
  5. Byers P. H. Ehlers-Danlos syndrome: recent advances and current understanding of the clinical and genetic heterogeneity. J Invest Dermatol. 1994 Nov;103(5 Suppl):47S–52S. doi: 10.1111/1523-1747.ep12399038. [DOI] [PubMed] [Google Scholar]
  6. Colige A., Beschin A., Samyn B., Goebels Y., Van Beeumen J., Nusgens B. V., Lapière C. M. Characterization and partial amino acid sequencing of a 107-kDa procollagen I N-proteinase purified by affinity chromatography on immobilized type XIV collagen. J Biol Chem. 1995 Jul 14;270(28):16724–16730. doi: 10.1074/jbc.270.28.16724. [DOI] [PubMed] [Google Scholar]
  7. Colige A., Li S. W., Sieron A. L., Nusgens B. V., Prockop D. J., Lapière C. M. cDNA cloning and expression of bovine procollagen I N-proteinase: a new member of the superfamily of zinc-metalloproteinases with binding sites for cells and other matrix components. Proc Natl Acad Sci U S A. 1997 Mar 18;94(6):2374–2379. doi: 10.1073/pnas.94.6.2374. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Colige A., Sieron A. L., Li S. W., Schwarze U., Petty E., Wertelecki W., Wilcox W., Krakow D., Cohn D. H., Reardon W. Human Ehlers-Danlos syndrome type VII C and bovine dermatosparaxis are caused by mutations in the procollagen I N-proteinase gene. Am J Hum Genet. 1999 Aug;65(2):308–317. doi: 10.1086/302504. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Evans J. P. Sperm disintegrins, egg integrins, and other cell adhesion molecules of mammalian gamete plasma membrane interactions. Front Biosci. 1999 Jan 15;4:D114–D131. doi: 10.2741/evans. [DOI] [PubMed] [Google Scholar]
  10. Georgiadis K. E., Hirohata S., Seldin M. F., Apte S. S. ADAM-TS8, a novel metalloprotease of the ADAM-TS family located on mouse chromosome 9 and human chromosome 11. Genomics. 1999 Dec 1;62(2):312–315. doi: 10.1006/geno.1999.6014. [DOI] [PubMed] [Google Scholar]
  11. Giunta C., Superti-Furga A., Spranger S., Cole W. G., Steinmann B. Ehlers-Danlos syndrome type VII: clinical features and molecular defects. J Bone Joint Surg Am. 1999 Feb;81(2):225–238. doi: 10.2106/00004623-199902000-00010. [DOI] [PubMed] [Google Scholar]
  12. Hasty P., Crist M., Grompe M., Bradley A. Efficiency of insertion versus replacement vector targeting varies at different chromosomal loci. Mol Cell Biol. 1994 Dec;14(12):8385–8390. doi: 10.1128/mcb.14.12.8385. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Hojima Y., McKenzie J. A., van der Rest M., Prockop D. J. Type I procollagen N-proteinase from chick embryo tendons. Purification of a new 500-kDa form of the enzyme and identification of the catalytically active polypeptides. J Biol Chem. 1989 Jul 5;264(19):11336–11345. [PubMed] [Google Scholar]
  14. Hojima Y., van der Rest M., Prockop D. J. Type I procollagen carboxyl-terminal proteinase from chick embryo tendons. Purification and characterization. J Biol Chem. 1985 Dec 15;260(29):15996–16003. [PubMed] [Google Scholar]
  15. Holmes D. F., Watson R. B., Steinmann B., Kadler K. E. Ehlers-Danlos syndrome type VIIB. Morphology of type I collagen fibrils formed in vivo and in vitro is determined by the conformation of the retained N-propeptide. J Biol Chem. 1993 Jul 25;268(21):15758–15765. [PubMed] [Google Scholar]
  16. Hooft van Huijsduijnen R. ADAM 20 and 21; two novel human testis-specific membrane metalloproteases with similarity to fertilin-alpha. Gene. 1998 Jan 12;206(2):273–282. doi: 10.1016/s0378-1119(97)00597-0. [DOI] [PubMed] [Google Scholar]
  17. Hulmes D. J., Kadler K. E., Mould A. P., Hojima Y., Holmes D. F., Cummings C., Chapman J. A., Prockop D. J. Pleomorphism in type I collagen fibrils produced by persistence of the procollagen N-propeptide. J Mol Biol. 1989 Nov 20;210(2):337–345. doi: 10.1016/0022-2836(89)90335-5. [DOI] [PubMed] [Google Scholar]
  18. Hurskainen T. L., Hirohata S., Seldin M. F., Apte S. S. ADAM-TS5, ADAM-TS6, and ADAM-TS7, novel members of a new family of zinc metalloproteases. General features and genomic distribution of the ADAM-TS family. J Biol Chem. 1999 Sep 3;274(36):25555–25563. doi: 10.1074/jbc.274.36.25555. [DOI] [PubMed] [Google Scholar]
  19. Khillan J. S., Bao Y. Preparation of animals with a high degree of chimerism by one-step coculture of embryonic stem cells and preimplantation embryos. Biotechniques. 1997 Mar;22(3):544–549. doi: 10.2144/97223rr06. [DOI] [PubMed] [Google Scholar]
  20. Király K., Hyttinen M. M., Lapveteläinen T., Elo M., Kiviranta I., Dobai J., Módis L., Helminen H. J., Arokoski J. P. Specimen preparation and quantification of collagen birefringence in unstained sections of articular cartilage using image analysis and polarizing light microscopy. Histochem J. 1997 Apr;29(4):317–327. doi: 10.1023/a:1020802631968. [DOI] [PubMed] [Google Scholar]
  21. Kuivaniemi H., Tromp G., Prockop D. J. Mutations in fibrillar collagens (types I, II, III, and XI), fibril-associated collagen (type IX), and network-forming collagen (type X) cause a spectrum of diseases of bone, cartilage, and blood vessels. Hum Mutat. 1997;9(4):300–315. doi: 10.1002/(SICI)1098-1004(1997)9:4<300::AID-HUMU2>3.0.CO;2-9. [DOI] [PubMed] [Google Scholar]
  22. Kuno K., Iizasa H., Ohno S., Matsushima K. The exon/intron organization and chromosomal mapping of the mouse ADAMTS-1 gene encoding an ADAM family protein with TSP motifs. Genomics. 1997 Dec 15;46(3):466–471. doi: 10.1006/geno.1997.5064. [DOI] [PubMed] [Google Scholar]
  23. Kuno K., Kanada N., Nakashima E., Fujiki F., Ichimura F., Matsushima K. Molecular cloning of a gene encoding a new type of metalloproteinase-disintegrin family protein with thrombospondin motifs as an inflammation associated gene. J Biol Chem. 1997 Jan 3;272(1):556–562. doi: 10.1074/jbc.272.1.556. [DOI] [PubMed] [Google Scholar]
  24. Lapière C. M., Lenaers A., Kohn L. D. Procollagen peptidase: an enzyme excising the coordination peptides of procollagen. Proc Natl Acad Sci U S A. 1971 Dec;68(12):3054–3058. doi: 10.1073/pnas.68.12.3054. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Li S. W., Khillan J., Prockop D. J. The complete cDNA coding sequence for the mouse pro alpha 1(I) chain of type I procollagen. Matrix Biol. 1995 Jul;14(7):593–595. doi: 10.1016/s0945-053x(05)80009-5. [DOI] [PubMed] [Google Scholar]
  26. Li S. W., Prockop D. J., Helminen H., Fässler R., Lapveteläinen T., Kiraly K., Peltarri A., Arokoski J., Lui H., Arita M. Transgenic mice with targeted inactivation of the Col2 alpha 1 gene for collagen II develop a skeleton with membranous and periosteal bone but no endochondral bone. Genes Dev. 1995 Nov 15;9(22):2821–2830. doi: 10.1101/gad.9.22.2821. [DOI] [PubMed] [Google Scholar]
  27. Långsjö T. K., Hyttinen M., Pelttari A., Kiraly K., Arokoski J., Helminen H. J. Electron microscopic stereological study of collagen fibrils in bovine articular cartilage: volume and surface densities are best obtained indirectly (from length densities and diameters) using isotropic uniform random sampling. J Anat. 1999 Aug;195(Pt 2):281–293. doi: 10.1046/j.1469-7580.1999.19520281.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Nusgens B. V., Verellen-Dumoulin C., Hermanns-Lê T., De Paepe A., Nuytinck L., Piérard G. E., Lapière C. M. Evidence for a relationship between Ehlers-Danlos type VII C in humans and bovine dermatosparaxis. Nat Genet. 1992 Jun;1(3):214–217. doi: 10.1038/ng0692-214. [DOI] [PubMed] [Google Scholar]
  29. Pereira R. F., Halford K. W., O'Hara M. D., Leeper D. B., Sokolov B. P., Pollard M. D., Bagasra O., Prockop D. J. Cultured adherent cells from marrow can serve as long-lasting precursor cells for bone, cartilage, and lung in irradiated mice. Proc Natl Acad Sci U S A. 1995 May 23;92(11):4857–4861. doi: 10.1073/pnas.92.11.4857. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Prockop D. J., Kivirikko K. I. Collagens: molecular biology, diseases, and potentials for therapy. Annu Rev Biochem. 1995;64:403–434. doi: 10.1146/annurev.bi.64.070195.002155. [DOI] [PubMed] [Google Scholar]
  31. Prockop D. J., Sieron A. L., Li S. W. Procollagen N-proteinase and procollagen C-proteinase. Two unusual metalloproteinases that are essential for procollagen processing probably have important roles in development and cell signaling. Matrix Biol. 1998 Feb;16(7):399–408. doi: 10.1016/s0945-053x(98)90013-0. [DOI] [PubMed] [Google Scholar]
  32. Reardon A., Sandell L., Jones C. J., McLeod D., Bishop P. N. Localization of pN-type IIA procollagen on adult bovine vitreous collagen fibrils. Matrix Biol. 2000 May;19(2):169–173. doi: 10.1016/s0945-053x(00)00058-5. [DOI] [PubMed] [Google Scholar]
  33. Shamsadin R., Adham I. M., Nayernia K., Heinlein U. A., Oberwinkler H., Engel W. Male mice deficient for germ-cell cyritestin are infertile. Biol Reprod. 1999 Dec;61(6):1445–1451. doi: 10.1095/biolreprod61.6.1445. [DOI] [PubMed] [Google Scholar]
  34. Tortorella M. D., Burn T. C., Pratta M. A., Abbaszade I., Hollis J. M., Liu R., Rosenfeld S. A., Copeland R. A., Decicco C. P., Wynn R. Purification and cloning of aggrecanase-1: a member of the ADAMTS family of proteins. Science. 1999 Jun 4;284(5420):1664–1666. doi: 10.1126/science.284.5420.1664. [DOI] [PubMed] [Google Scholar]
  35. Vandenberg P., Khillan J. S., Prockop D. J., Helminen H., Kontusaari S., Ala-Kokko L. Expression of a partially deleted gene of human type II procollagen (COL2A1) in transgenic mice produces a chondrodysplasia. Proc Natl Acad Sci U S A. 1991 Sep 1;88(17):7640–7644. doi: 10.1073/pnas.88.17.7640. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Vázquez F., Hastings G., Ortega M. A., Lane T. F., Oikemus S., Lombardo M., Iruela-Arispe M. L. METH-1, a human ortholog of ADAMTS-1, and METH-2 are members of a new family of proteins with angio-inhibitory activity. J Biol Chem. 1999 Aug 13;274(33):23349–23357. doi: 10.1074/jbc.274.33.23349. [DOI] [PubMed] [Google Scholar]
  37. Watson R. B., Holmes D. F., Graham H. K., Nusgens B. V., Kadler K. E. Surface located procollagen N-propeptides on dermatosparactic collagen fibrils are not cleaved by procollagen N-proteinase and do not inhibit binding of decorin to the fibril surface. J Mol Biol. 1998 Apr 24;278(1):195–204. doi: 10.1006/jmbi.1998.1680. [DOI] [PubMed] [Google Scholar]
  38. Yuan R., Primakoff P., Myles D. G. A role for the disintegrin domain of cyritestin, a sperm surface protein belonging to the ADAM family, in mouse sperm-egg plasma membrane adhesion and fusion. J Cell Biol. 1997 Apr 7;137(1):105–112. doi: 10.1083/jcb.137.1.105. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Zhu G. Z., Lin Y., Myles D. G., Primakoff P. Identification of four novel ADAMs with potential roles in spermatogenesis and fertilization. Gene. 1999 Jul 8;234(2):227–237. doi: 10.1016/s0378-1119(99)00208-5. [DOI] [PubMed] [Google Scholar]

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