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.
Delorme
V G
VG
Department of Plant Sciences, University of Oxford, South Parks Road, Oxford OX1 3RB, United Kingdom.
McCabe
P F
PF
Kim
D J
DJ
Leaver
C J
CJ
D020816:Amino Acid Motifs; D000595:Amino Acid Sequence; D017209:Apoptosis / Q000235:genetics*; D001483:Base Sequence; D015152:Blotting, Northern; D015139:Blotting, Southern; D020134:Catalytic Domain; D016922:Cellular Senescence / Q000235:genetics*; D003094:Collagen / Q000378:metabolism; D017364:Collagenases / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D018548:Cotyledon / Q000201:enzymology / Q000235:genetics / Q000378:metabolism; D018553:Cucumis sativus / Q000201:enzymology* / Q000235:genetics / Q000378:metabolism; D053938:DNA Fragmentation; D005109:Extracellular Matrix / Q000201:enzymology; D020782:Matrix Metalloproteinases / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism; D008969:Molecular Sequence Data; D010940:Plant Proteins; D016133:Polymerase Chain Reaction; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA; D015032:Zinc / Q000378:metabolism
D010940:Plant Proteins; D003094:Collagen; D017364:Collagenases; C414310:Cs1-MMP protein, Cucumis sativus; D020782:Matrix Metalloproteinases; D015032:Zinc