Logo of jcellbiolHomeThe Rockefeller University PressEditorsContactInstructions for AuthorsThis issue
J Cell Biol. 1995 Jul 1; 130(1): 227–237.
PMCID: PMC2120510

A targeted mutation at the known collagenase cleavage site in mouse type I collagen impairs tissue remodeling


Degradation of type I collagen, the most abundant collagen, is initiated by collagenase cleavage at a highly conserved site between Gly775 and Ile776 of the alpha 1 (I) chain. Mutations at or around this site render type I collagen resistant to collagenase digestion in vitro. We show here that mice carrying a collagenase-resistant mutant Col1a-1 transgene die late in embryo-genesis, ascribable to overexpression of the transgene, since the same mutation introduced into the endogenous Col1a-1 gene by gene targeting permitted normal development of mutant mice to young adulthood. With increasing age, animals carrying the targeted mutation developed marked fibrosis of the dermis similar to that in human scleroderma. Postpartum involution of the uterus in the mutant mice was also impaired, with persistence of collagenous nodules in the uterine wall. Although type I collagen from the homozygous mutant mice was resistant to cleavage by human or rat fibroblast collagenases at the helical site, only the rat collagenase cleaved collagen trimers at an additional, novel site in the nonhelical N-telopeptide domain. Our results suggest that cleavage by murine collagenase at the N-telopeptide site could account for resorption of type I collagen during embryonic and early adult life. During intense collagen resorption, however, such as in the immediate postpartum uterus and in the dermis later in life, cleavage at the helical site is essential for normal collagen turnover. Thus, type I collagen is degraded by at least two differentially controlled mechanisms involving collagenases with distinct, but overlapping, substrate specificities.

Full Text

The Full Text of this article is available as a PDF (6.4M).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.
  • Aimes RT, Quigley JP. Matrix metalloproteinase-2 is an interstitial collagenase. Inhibitor-free enzyme catalyzes the cleavage of collagen fibrils and soluble native type I collagen generating the specific 3/4- and 1/4-length fragments. J Biol Chem. 1995 Mar 17;270(11):5872–5876. [PubMed]
  • Alexander CM, Werb Z. Proteinases and extracellular matrix remodeling. Curr Opin Cell Biol. 1989 Oct;1(5):974–982. [PubMed]
  • Bernfield M, Banerjee SD, Koda JE, Rapraeger AC. Remodelling of the basement membrane: morphogenesis and maturation. Ciba Found Symp. 1984;108:179–196. [PubMed]
  • Birkedal-Hansen H. Catabolism and turnover of collagens: collagenases. Methods Enzymol. 1987;144:140–171. [PubMed]
  • Birkedal-Hansen H, Moore WG, Bodden MK, Windsor LJ, Birkedal-Hansen B, DeCarlo A, Engler JA. Matrix metalloproteinases: a review. Crit Rev Oral Biol Med. 1993;4(2):197–250. [PubMed]
  • Bonadio J, Saunders TL, Tsai E, Goldstein SA, Morris-Wiman J, Brinkley L, Dolan DF, Altschuler RA, Hawkins JE, Jr, Bateman JF, et al. Transgenic mouse model of the mild dominant form of osteogenesis imperfecta. Proc Natl Acad Sci U S A. 1990 Sep;87(18):7145–7149. [PMC free article] [PubMed]
  • Byers PH, Steiner RD. Osteogenesis imperfecta. Annu Rev Med. 1992;43:269–282. [PubMed]
  • Dayer JM, Stephenson ML, Schmidt E, Karge W, Krane SM. Purification of a factor from human blood monocyte-macrophages which stimulates the production of collagenase and prostaglandin E2 by cells cultured from rheumatoid synovial tissues. FEBS Lett. 1981 Feb 23;124(2):253–253. [PubMed]
  • Freije JM, Díez-Itza I, Balbín M, Sánchez LM, Blasco R, Tolivia J, López-Otín C. Molecular cloning and expression of collagenase-3, a novel human matrix metalloproteinase produced by breast carcinomas. J Biol Chem. 1994 Jun 17;269(24):16766–16773. [PubMed]
  • Goldberg GI, Wilhelm SM, Kronberger A, Bauer EA, Grant GA, Eisen AZ. Human fibroblast collagenase. Complete primary structure and homology to an oncogene transformation-induced rat protein. J Biol Chem. 1986 May 15;261(14):6600–6605. [PubMed]
  • Goldring MB, Krane SM. Modulation by recombinant interleukin 1 of synthesis of types I and III collagens and associated procollagen mRNA levels in cultured human cells. J Biol Chem. 1987 Dec 5;262(34):16724–16729. [PubMed]
  • HARKNESS ML, HARKNESS RD. The distribution of the growth of collagen in the uterus of the pregnant rat. J Physiol. 1956 Jun 28;132(3):492–501. [PMC free article] [PubMed]
  • HARKNESS RD, MORALEE BE. The time-course and route of loss of collagen from the rat's uterus during post-partum involution. J Physiol. 1956 Jun 28;132(3):502–508. [PMC free article] [PubMed]
  • Hartung S, Jaenisch R, Breindl M. Retrovirus insertion inactivates mouse alpha 1(I) collagen gene by blocking initiation of transcription. Nature. 320(6060):365–367. [PubMed]
  • Hasty KA, Pourmotabbed TF, Goldberg GI, Thompson JP, Spinella DG, Stevens RM, Mainardi CL. Human neutrophil collagenase. A distinct gene product with homology to other matrix metalloproteinases. J Biol Chem. 1990 Jul 15;265(20):11421–11424. [PubMed]
  • Hasty P, Ramírez-Solis R, Krumlauf R, Bradley A. Introduction of a subtle mutation into the Hox-2.6 locus in embryonic stem cells. Nature. 1991 Mar 21;350(6315):243–246. [PubMed]
  • Henriet P, Rousseau GG, Eeckhout Y. Cloning and sequencing of mouse collagenase cDNA. Divergence of mouse and rat collagenases from the other mammalian collagenases. FEBS Lett. 1992 Sep 28;310(2):175–178. [PubMed]
  • Herskowitz I. Functional inactivation of genes by dominant negative mutations. Nature. 1987 Sep 17;329(6136):219–222. [PubMed]
  • Jacenko O, LuValle PA, Olsen BR. Spondylometaphyseal dysplasia in mice carrying a dominant negative mutation in a matrix protein specific for cartilage-to-bone transition. Nature. 1993 Sep 2;365(6441):56–61. [PubMed]
  • Jeffrey JJ. Collagen and collagenase: pregnancy and parturition. Semin Perinatol. 1991 Apr;15(2):118–126. [PubMed]
  • Laird PW, Zijderveld A, Linders K, Rudnicki MA, Jaenisch R, Berns A. Simplified mammalian DNA isolation procedure. Nucleic Acids Res. 1991 Aug 11;19(15):4293–4293. [PMC free article] [PubMed]
  • Leivo I, Vaheri A, Timpl R, Wartiovaara J. Appearance and distribution of collagens and laminin in the early mouse embryo. Dev Biol. 1980 Apr;76(1):100–114. [PubMed]
  • Li E, Bestor TH, Jaenisch R. Targeted mutation of the DNA methyltransferase gene results in embryonic lethality. Cell. 1992 Jun 12;69(6):915–926. [PubMed]
  • Löhler J, Timpl R, Jaenisch R. Embryonic lethal mutation in mouse collagen I gene causes rupture of blood vessels and is associated with erythropoietic and mesenchymal cell death. Cell. 1984 Sep;38(2):597–607. [PubMed]
  • Matsudaira P. Limited N-terminal sequence analysis. Methods Enzymol. 1990;182:602–613. [PubMed]
  • Mauch C, Eckes B, Hunzelmann N, Oono T, Kozlowska E, Krieg T. Control of fibrosis in systemic scleroderma. J Invest Dermatol. 1993 Jan;100(1):92S–96S. [PubMed]
  • MORRIONE TG, SEIFTER S. Alteration in the collagen content of the human uterus during pregnancy and post partum involution. J Exp Med. 1962 Feb 1;115:357–365. [PMC free article] [PubMed]
  • Murphy G, Atkinson S, Ward R, Gavrilovic J, Reynolds JJ. The role of plasminogen activators in the regulation of connective tissue metalloproteinases. Ann N Y Acad Sci. 1992 Dec 4;667:1–12. [PubMed]
  • Murphy G, Docherty AJ. The matrix metalloproteinases and their inhibitors. Am J Respir Cell Mol Biol. 1992 Aug;7(2):120–125. [PubMed]
  • Nagase H, Jackson RC, Brinckerhoff CE, Vater CA, Harris ED., Jr A precursor form of latent collagenase produced in a cell-free system with mRNA from rabbit synovial cells. J Biol Chem. 1981 Dec 10;256(23):11951–11954. [PubMed]
  • Penn PE, Jiang DZ, Fei RG, Sitnicka E, Wolf NS. Dissecting the hematopoietic microenvironment. IX. Further characterization of murine bone marrow stromal cells. Blood. 1993 Mar 1;81(5):1205–1213. [PubMed]
  • Pereira R, Halford K, Sokolov BP, Khillan JS, Prockop DJ. Phenotypic variability and incomplete penetrance of spontaneous fractures in an inbred strain of transgenic mice expressing a mutated collagen gene (COL1A1). J Clin Invest. 1994 Apr;93(4):1765–1769. [PMC free article] [PubMed]
  • Quinn CO, Scott DK, Brinckerhoff CE, Matrisian LM, Jeffrey JJ, Partridge NC. Rat collagenase. Cloning, amino acid sequence comparison, and parathyroid hormone regulation in osteoblastic cells. J Biol Chem. 1990 Dec 25;265(36):22342–22347. [PubMed]
  • Rintala M, Metsäranta M, Garofalo S, de Crombrugghe B, Vuorio E, Rönning O. Abnormal craniofacial morphology and cartilage structure in transgenic mice harboring a Gly --> Cys mutation in the cartilage-specific type II collagen gene. J Craniofac Genet Dev Biol. 1993 Jul-Sep;13(3):137–146. [PubMed]
  • Roswit WT, Halme J, Jeffrey JJ. Purification and properties of rat uterine procollagenase. Arch Biochem Biophys. 1983 Aug;225(1):285–295. [PubMed]
  • Shimizu K, Maekawa K. Collagen degradation in the mouse uterus during postpartum involution: extracellular pathway. Acta Anat (Basel) 1983;117(3):257–260. [PubMed]
  • Shimizu K, Hokano M. Removal of collagen bundles in murine uterus during postpartum involution. Anat Rec. 1988 Feb;220(2):138–142. [PubMed]
  • Sottrup-Jensen L, Birkedal-Hansen H. Human fibroblast collagenase-alpha-macroglobulin interactions. Localization of cleavage sites in the bait regions of five mammalian alpha-macroglobulins. J Biol Chem. 1989 Jan 5;264(1):393–401. [PubMed]
  • Stacey A, Bateman J, Choi T, Mascara T, Cole W, Jaenisch R. Perinatal lethal osteogenesis imperfecta in transgenic mice bearing an engineered mutant pro-alpha 1(I) collagen gene. Nature. 1988 Mar 10;332(6160):131–136. [PubMed]
  • Sympson CJ, Talhouk RS, Alexander CM, Chin JR, Clift SM, Bissell MJ, Werb Z. Targeted expression of stromelysin-1 in mammary gland provides evidence for a role of proteinases in branching morphogenesis and the requirement for an intact basement membrane for tissue-specific gene expression. J Cell Biol. 1994 May;125(3):681–693. [PMC free article] [PubMed]
  • Vandenberg P, Khillan JS, Prockop DJ, 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. [PMC free article] [PubMed]
  • Vuorio E, de Crombrugghe B. The family of collagen genes. Annu Rev Biochem. 1990;59:837–872. [PubMed]
  • Welgus HG, Kobayashi DK, Jeffrey JJ. The collagen substrate specificity of rat uterus collagenase. J Biol Chem. 1983 Dec 10;258(23):14162–14165. [PubMed]
  • Whitham SE, Murphy G, Angel P, Rahmsdorf HJ, Smith BJ, Lyons A, Harris TJ, Reynolds JJ, Herrlich P, Docherty AJ. Comparison of human stromelysin and collagenase by cloning and sequence analysis. Biochem J. 1986 Dec 15;240(3):913–916. [PMC free article] [PubMed]
  • Wilkie TM, Brinster RL, Palmiter RD. Germline and somatic mosaicism in transgenic mice. Dev Biol. 1986 Nov;118(1):9–18. [PubMed]
  • Woessner JF., Jr Total, latent and active collagenase during the course of post-partum involution of the rat uterus. Effect of oestradiol. Biochem J. 1979 Apr 15;180(1):95–102. [PMC free article] [PubMed]
  • Woessner JF., Jr Matrix metalloproteinases and their inhibitors in connective tissue remodeling. FASEB J. 1991 May;5(8):2145–2154. [PubMed]
  • Wu H, Byrne MH, Stacey A, Goldring MB, Birkhead JR, Jaenisch R, Krane SM. Generation of collagenase-resistant collagen by site-directed mutagenesis of murine pro alpha 1(I) collagen gene. Proc Natl Acad Sci U S A. 1990 Aug;87(15):5888–5892. [PMC free article] [PubMed]
  • Wu H, Bateman JF, Schnieke A, Sharpe A, Barker D, Mascara T, Eyre D, Bruns R, Krimpenfort P, Berns A, et al. Human-mouse interspecies collagen I heterotrimer is functional during embryonic development of Mov13 mutant mouse embryos. Mol Cell Biol. 1990 Apr;10(4):1452–1460. [PMC free article] [PubMed]
  • Wu H, Liu X, Jaenisch R. Double replacement: strategy for efficient introduction of subtle mutations into the murine Col1a-1 gene by homologous recombination in embryonic stem cells. Proc Natl Acad Sci U S A. 1994 Mar 29;91(7):2819–2823. [PMC free article] [PubMed]
  • Wu JJ, Lark MW, Chun LE, Eyre DR. Sites of stromelysin cleavage in collagen types II, IX, X, and XI of cartilage. J Biol Chem. 1991 Mar 25;266(9):5625–5628. [PubMed]

Articles from The Journal of Cell Biology are provided here courtesy of The Rockefeller University Press


Save items

Related citations in PubMed

See reviews...See all...

Cited by other articles in PMC

See all...


  • Gene
    Gene records that cite the current articles. Citations in Gene are added manually by NCBI or imported from outside public resources.
  • GEO Profiles
    GEO Profiles
    Gene Expression Omnibus (GEO) Profiles of molecular abundance data. The current articles are references on the Gene record associated with the GEO profile.
  • HomoloGene
    HomoloGene clusters of homologous genes and sequences that cite the current articles. These are references on the Gene and sequence records in the HomoloGene entry.
  • MedGen
    Related information in MedGen
  • Pathways + GO
    Pathways + GO
    Pathways and biological systems (BioSystems) that cite the current articles. Citations are from the BioSystems source databases (KEGG and BioCyc).
  • PubMed
    PubMed citations for these articles
  • Substance
    PubChem chemical substance records that cite the current articles. These references are taken from those provided on submitted PubChem chemical substance records.

Recent Activity

Your browsing activity is empty.

Activity recording is turned off.

Turn recording back on

See more...