A matrix metalloproteinase gene is expressed at the boundary of senescence and programmed cell death in cucumber.

Cell-cell and extracellular cell matrix (ECM) interactions provide cells with information essential for controlling morphogenesis, cell-fate specification, and cell death. In animals, one of the major groups of enzymes that degrade the ECM is the matrix metalloproteinases (MMPs). Here, we report the characterization of the cucumber (Cucumis sativus L. cv Marketmore) Cs1-MMP gene encoding such an enzyme likely to play a role in plant ECM degradation. Cs1-MMP has all the hallmark motif characteristics of animal MMPs and is a pre-pro-enzyme having a signal peptide, propeptide, and zinc-binding catalytic domains. Cs1-MMP also displays functional similarities with animal MMPs. For example, it has a collagenase-like activity that can cleave synthetic peptides and type-I collagen, a major component of animal ECM. Cs1-MMP activity is completely inhibited by a hydroxamate-based inhibitor that binds at the active site of MMPs in a stereospecific manner. The Cs1-MMP gene is expressed de novo at the end stage of developmental senescence, prior to the appearance of DNA laddering in cucumber cotyledons leaf discs and male flowers. As the steady-state level of Cs1-MMP mRNA peaks late in senescence and the pro-enzyme must undergo maturation and activation, the protease is probably not involved in nutrient remobilization during senescence but may have another function. The physiological substrates for Cs1-MMP remain to be determined, but the enzyme represents a good candidate for plant ECM degradation and may be involved in programmed cell death (PCD). Our results suggest that PCD occurs only at the culmination of the senescence program or that the processes are distinct with PCD being triggered at the end of senescence.

[1]  G. Murphy,et al.  Gelatinases A and B. , 2002, Methods in enzymology.

[2]  J. Sambrook,et al.  Molecular Cloning: A Laboratory Manual , 2001 .

[3]  G. Murphy,et al.  Matrix Metalloproteinase Homologues from Arabidopsis thaliana , 1999, The Journal of Biological Chemistry.

[4]  Jones,et al.  Tracheary element differentiation uses a novel mechanism coordinating programmed cell death and secondary cell wall synthesis , 1999, Plant physiology.

[5]  E. Lam,et al.  Caspases and programmed cell death in the hypersensitive response of plants to pathogens , 1998, Current Biology.

[6]  A. Danon,et al.  UV‐C radiation induces apoptotic‐like changes in Arabidopsis thaliana , 1998, FEBS letters.

[7]  E. Lam,et al.  Caspases and programmed cell death in the hypersensitive response of plants to pathogens , 1998, Current Biology.

[8]  Gabriele Bergers,et al.  MMP-9/Gelatinase B Is a Key Regulator of Growth Plate Angiogenesis and Apoptosis of Hypertrophic Chondrocytes , 1998, Cell.

[9]  H. Hayashi,et al.  Two vascular apoptosis-inducing proteins from snake venom are members of the metalloprotease/disintegrin family. , 1998, European journal of biochemistry.

[10]  R. Timpl,et al.  Membrane-type matrix metalloproteinases 1 and 2 exhibit broad-spectrum proteolytic capacities comparable to many matrix metalloproteinases. , 1997, European journal of biochemistry.

[11]  Z. Werb.  ECM and Cell Surface Proteolysis: Regulating Cellular Ecology , 1997, Cell.

[12]  C. Keen,et al.  Cytoplasmic-nuclear male sterility in pearl millet: comparative RFLP and transcript analyses of isonuclear male-sterile lines , 1997, Theoretical and Applied Genetics.

[13]  N. Tapon,et al.  A programmed cell death pathway activated in carrot cells cultured at low cell density , 1997 .

[14]  G. Butler,et al.  Membrane-type-2 matrix metalloproteinase can initiate the processing of progelatinase A and is regulated by the tissue inhibitors of metalloproteinases. , 1997, European journal of biochemistry.

[15]  D. -. Kim,et al.  A cDNA encoding a putative SPF1-type DNA-binding protein from cucumber. , 1997, Gene.

[16]  P. Angel,et al.  The catalytic domain of activated collagenase I (MMP-1) is absolutely required for interaction with its specific inhibitor, tissue inhibitor of metalloproteinases-1 (TIMP-1). , 1997, European journal of biochemistry.

[17]  R. Amasino,et al.  Making Sense of Senescence (Molecular Genetic Regulation and Manipulation of Leaf Senescence) , 1997, Plant physiology.

[18]  E. Lam,et al.  Inhibition of Programmed Cell Death in Tobacco Plants during a Pathogen-Induced Hypersensitive Response at Low Oxygen Pressure. , 1996, The Plant cell.

[19]  C. Lamb,et al.  Calcium-mediated apoptosis in a plant hypersensitive disease resistance response , 1996, Current Biology.

[20]  D. Ryerson,et al.  Cleavage of Nuclear DNA into Oligonucleosomal Fragments during Cell Death Induced by Fungal Infection or by Abiotic Treatments. , 1996, The Plant cell.

[21]  C. López-Otín,et al.  Biochemical Characterization of Human Collagenase-3 (*) , 1996, The Journal of Biological Chemistry.

[22]  P. Reinemer,et al.  The metzincins — Topological and sequential relations between the astacins, adamalysins, serralysins, and matrixins (collagenases) define a super family of zinc‐peptidases , 1995, Protein science : a publication of the Protein Society.

[23]  A. Hassell,et al.  Crystal structures of recombinant 19-kDa human fibroblast collagenase complexed to itself. , 1995, Biochemistry.

[24]  J. Foidart,et al.  Matrix metalloproteinase family. , 1995, Cell biology international.

[25]  N. Hooper,et al.  Families of zinc metalloproteases , 1994, FEBS letters.

[26]  M. Cockett,et al.  Assessment of the role of the fibronectin-like domain of gelatinase A by analysis of a deletion mutant. , 1994, The Journal of biological chemistry.

[27]  Catherine M Smart,et al.  Gene expression during leaf senescence. , 1994, The New phytologist.

[28]  A M Hassell,et al.  Structure of the catalytic domain of fibroblast collagenase complexed with an inhibitor. , 1994, Science.

[29]  W. Bode,et al.  Astacins, serralysins, snake venom and matrix metalloproteinases exhibit identical zinc‐binding environments (HEXXHXXGXXH and Met‐turn) and topologies and should be grouped into a common family, the ‘metzincins’ , 1993, FEBS letters.

[30]  J. Marrison,et al.  Co‐immunolocalization of topoisomerase II and chloroplast DNA in developing, dividing and mature wheat chloroplasts , 1992 .

[31]  R. Anderegg,et al.  Sequencing and characterization of the soybean leaf metalloproteinase : structural and functional similarity to the matrix metalloproteinase family. , 1992, Plant physiology.

[32]  H. Fukuzawa,et al.  Primary structure and expression of a gamete lytic enzyme in Chlamydomonas reinhardtii: similarity of functional domains to matrix metalloproteases. , 1992, Proceedings of the National Academy of Sciences of the United States of America.

[33]  I. Graham,et al.  Induction of Malate Synthase Gene Expression in Senescent and Detached Organs of Cucumber. , 1992, The Plant cell.

[34]  G Murphy,et al.  A novel coumarin‐labelled peptide for sensitive continuous assays of the matrix metalloproteinases , 1992, FEBS letters.

[35]  J. Graham,et al.  Purification and Developmental Analysis of a Metalloendoproteinase from the Leaves of Glycine max. , 1991, Plant physiology.

[36]  H. Birkedal‐Hansen,et al.  The cysteine switch: a principle of regulation of metalloproteinase activity with potential applicability to the entire matrix metalloproteinase gene family. , 1990, Proceedings of the National Academy of Sciences of the United States of America.

[37]  M. Frohman,et al.  Rapid production of full-length cDNAs from rare transcripts: amplification using a single gene-specific oligonucleotide primer. , 1988, Proceedings of the National Academy of Sciences of the United States of America.

[38]  J. Draper.  Plant Genetic Transformation and Gene Expression: A Laboratory Manual , 1988 .

[39]  S. Imam,et al.  The Chlamydomonas cell wall degrading enzyme, lysin, acts on two substrates within the framework of the wall , 1988, The Journal of cell biology.

[40]  H. Riezman,et al.  Regulation of Glyoxysomal Enzymes during Germination of Cucumber: I. Developmental Changes in Cotyledonary Protein, RNA, and Enzyme Activities during Germination. , 1978, Plant physiology.

[41]  P. Matile,et al.  Function of Lysosomes and Lysosomal Enzymes in the Senescing Corolla of the Morning Glory (Ipomoea purpurea) , 1971 .

[42]  U. K. Laemmli,et al.  Cleavage of Structural Proteins during the Assembly of the Head of Bacteriophage T4 , 1970, Nature.

[43]  H. E. Wart,et al.  Substrate Specificity and Mechanisms of Substrate Recognition of the Matrix Metalloproteinases , 1998 .

[44]  A. Granell,et al.  DNA fragmentation is regulated by ethylene during carpel senescence in Pisum sativum , 1997 .

[45]  J. Guiamet,et al.  Senescence mechanisms : Special section: hormones, regulating and signaling substances in plant growth and development. Part 2 , 1997 .

[46]  A. H. Drummond,et al.  Recent advances in matrix metalloproteinase inhibitor research , 1996 .

[47]  P. Schopfer,et al.  Arabinogalactan proteins in maize coleoptiles: developmental relationship to cell death during xylem differentiation but not to extension growth. , 1995, The Plant journal : for cell and molecular biology.

[48]  H. Birkedal‐Hansen,et al.  Multiple modes of activation of latent human fibroblast collagenase: evidence for the role of a Cys73 active-site zinc complex in latency and a "cysteine switch" mechanism for activation. , 1990, Proceedings of the National Academy of Sciences of the United States of America.

[49]  D. Arnon.  COPPER ENZYMES IN ISOLATED CHLOROPLASTS. POLYPHENOLOXIDASE IN BETA VULGARIS. , 1949, Plant physiology.