Degradomic and yeast 2-hybrid inactive catalytic domain substrate trapping identifies new membrane-type 1 matrix metalloproteinase (MMP14) substrates: CCN3 (Nov) and CCN5 (WISP2).
; ; Sounni, Nor Eddine et al
in Matrix Biology (2016)
Members of the CCN family of matricellular proteins are cytokines linking cells to the extracellular matrix. We report that CCN3 (Nov) and CCN5 (WISP2) are novel substrates of MMP14 (membrane-type 1 ... [more ▼]
Members of the CCN family of matricellular proteins are cytokines linking cells to the extracellular matrix. We report that CCN3 (Nov) and CCN5 (WISP2) are novel substrates of MMP14 (membrane-type 1-matrix metalloproteinase, MT1-MMP) that we identified using MMP14 "inactive catalytic domain capture" (ICDC) as a yeast two-hybrid protease substrate trapping platform in parallel with degradomics mass spectrometry screens for MMP14 substrates. CCN3 and CCN5, previously unknown substrates of MMPs, were biochemically validated as substrates of MMP14 and other MMPs in vitro-CCN5 was processed in the variable region by MMP14 and MMP2, as well as by MMP1, 3, 7, 8, 9 and 15. CCN1, 2 and 3 are proangiogenic factors yet we found novel opposing activity of CCN5 that was potently antiangiogenic in an aortic ring vessel outgrowth model. MMP14, a known regulator of angiogenesis, cleaved CCN5 and abrogated the angiostatic activity. CCN3 was also processed in the variable region by MMP14 and MMP2, and by MMP1, 8 and 9. In addition to the previously reported cleavages of CCN1 and CCN2 by several MMPs we found that MMPs 8, 9, and 1 process CCN1, and MMP8 and MMP9 also process CCN2. Thus, our study reveals additional and pervasive family-wide processing of CCN matricellular proteins/cytokines by MMPs. Furthermore, CCN5 cleavage by proangiogenic MMPs results in removal of an angiogenic brake held by CCN5. This highlights the importance of thorough dissection of MMP substrates that is needed to reveal higher-level control mechanisms beyond type IV collagen and other extracellular matrix protein remodelling in angiogenesis. SUMMARY: We find CCN family member cleavage by MMPs is more pervasive than previously reported and includes CCN3 (Nov) and CCN5 (WISP2). CCN5 is a novel antiangiogenic factor, whose function is abrogated by proangiogenic MMP cleavage. By processing CCN proteins, MMPs regulate cell responses angiogenesis in connective tissues. [less ▲]Detailed reference viewed: 13 (1 ULg)
The procollagen N-proteinases ADAMTS2, 3 and 14 in pathophysiology.
Bekhouche, Mourad ; Colige, Alain
in Matrix Biology (2015), 44-46Detailed reference viewed: 19 (1 ULg)
Osterix/Sp7 regulates biomineralization of otoliths and bone in medaka (Oryzias latipes).
Renn, Jörg ;
in Matrix Biology (2014), 34
Osterix/Sp7 is a zinc finger transcription factor and critical regulator of osteoblast differentiation,maturation and activity. Osterix expression has also been described in non-skeletal tissues but ... [more ▼]
Osterix/Sp7 is a zinc finger transcription factor and critical regulator of osteoblast differentiation,maturation and activity. Osterix expression has also been described in non-skeletal tissues but functional analyses are lacking. In the present study, we show that in the teleost model medaka, osterix is present as two alternatively spliced transcripts, osx_tv1 and osx_tv2. Knock-down of osx_tv1 and/or osx_tv2 results in mineralization loss in early intramembranous bones while cartilage formation ismostly unaffected. Formation of the parasphenoid, the earliestmineralized bone in themedaka skeleton, is impaired and fails to recover at later stages. Ossification of later bones, such as the operculum and cleithrum, is delayed but recovers during further development. In the axial skeleton, formation of the neural arches and centra is strongly delayed. In vivo analyses using osterix:nlGFP and osteocalcin:GFP transgenic medaka and whole mount in situ hybridization suggest that bone defects observed after knock-down of osterix are caused by a delay of osteoblast maturation and activity. Furthermore, we analyzed expression profile and function of osterix during ear and otolith formation. We show that osterix is expressed in otic placodes at the otic vesicle stage and that its knock-down results in a loss of otoliths. Taken together, we show that osterix is required for bone formation in a teleost fish and that its important regulatory functions are conserved between teleosts and mammals. Furthermore, we provide the first functional evidence for a role of Osterix in a non-skeletal tissue, i.e. the otoliths. [less ▲]Detailed reference viewed: 57 (2 ULg)
Membrane type-1 matrix metalloproteinase and TIMP-2 in tumor angiogenesis
Sounni, Nor Eddine ; ; Foidart, Jean-Michel et al
in Matrix Biology (2003), 22(1), 55-61
The matrix metalloproteinases (MMPs) constitute a multigene family of over 23 secreted and cell-surface associated enzymes that cleave or degrade various pericellular substrates. In addition to virtually ... [more ▼]
The matrix metalloproteinases (MMPs) constitute a multigene family of over 23 secreted and cell-surface associated enzymes that cleave or degrade various pericellular substrates. In addition to virtually all extracellular matrix (ECM) compounds, their targets include other proteinases, chemotactic molecules, latent growth factors, growth factor-binding proteins and cell surface molecules. The MMP activity is controlled by the physiological tissue inhibitors of MMPs (TIMPs). There is much evidence that MMPs and their inhibitors play a key role during extracellular remodeling in physiological situations and in cancer progression. They have other functions that promoting tumor invasion. Indeed, they regulate early stages of tumor progression such as tumor growth and angiogenesis. Membrane type MMPs (MT-MMPs) constitute a new subset of cell surface-associated MMPs. The present review will focus on MT1-MMP which plays a major role at least, in the ECM remodeling, directly by degrading several of its components, and indirectly by activating pro-MMP2. As our knowledge on the field of MT1-MMP biology has grown, the unforeseen complexities of this enzyme and its interaction with its inhibitor TIMP-2 have emerged, often revealing unexpected mechanisms of action. (C) 2003 Elsevier Science B.V/Intemational Society of Matrix Biology. All rights reserved. [less ▲]Detailed reference viewed: 46 (3 ULg)
Distinct pathways in the over-expression of matrix metalloproteinases in human fibroblasts by relaxation of mechanical tension.
Lambert, Charles ; Colige, Alain ; Munaut, Carine et al
in Matrix Biology (2001), 20(7), 397-408
The aim of the work was to analyze, on a comparative basis, the signaling pathways operating in the regulation of a panel of matrix metalloproteinases (MMP) expressed by human dermal fibroblasts submitted ... [more ▼]
The aim of the work was to analyze, on a comparative basis, the signaling pathways operating in the regulation of a panel of matrix metalloproteinases (MMP) expressed by human dermal fibroblasts submitted to mechanical stress relaxation by cytochalasin D (CD) and in a retracting collagen gel (RCG). The mRNA steady-state level of MMPs was measured by a quantitative RT-PCR procedure using a synthetic RNA as internal standard. In monolayer, most MMPs were barely detected, except MMP-2. Disruption of the actin stress fibers by CD induced a moderate increase of MMP-2 mRNA and a much larger stimulation of MMP-3, -9, -13 and -14 mRNAs. In RCG, a significant up-regulation of these MMPs was also observed although to a lower extent than in CD-treated monolayers. Among the investigated MMPs, the MMP-8 and -11 were not reproducibly detected. MMP-2 was processed to its active form both by CD and in RCG. The CD-induced up-regulation of gene expression was largely repressed by blocking protein synthesis by cycloheximide for all the MMPs, by inhibiting the tyrosine-kinases of the src family by herbimycin A for all MMPs, except MMP-2, and by inhibiting the TPA-inducible PKC isoforms by bisindoyl maleimide for all MMPs, except MMP-14. The up-regulation induced by stress relaxation in RCG was protein synthesis-dependent for MMP-2 and MMP-13, tyrosine kinases-dependent for MMP-3 and MMP-13, as previously described for MMP-1. Inhibiting TPA-inducible PKC did not affect any MMP in RCG except MMP-13, which was strongly induced. The processing of MMP-2 was tyrosine kinases-dependent but PKC-independent. Inhibitors of the ERK1,2 and p38 MAP kinases pathways diversely affected the MMPs expression. Inhibiting the Rho-kinase activity by Y-27632 was inactive. These results point to the potent regulation operated by the status of the cytoskeleton on the cell phenotype, and to distinct regulatory pathways involved in the control of different MMPs expression. [less ▲]Detailed reference viewed: 42 (2 ULg)
Analysis of coding sequences for tissue inhibitor of metalloproteinases 1 (TIMP1) and 2 (TIMP2) in patients with aneurysms.
Wang, Xingjun ; ; et al
in Matrix Biology (1999), 18(2), 121-4
Aneurysms are characterized by dilation, i.e. expansion and thinning of all the arterial wall layers, which is accompanied by remodeling of the connective tissue. Genes involved in the regulation of ... [more ▼]
Aneurysms are characterized by dilation, i.e. expansion and thinning of all the arterial wall layers, which is accompanied by remodeling of the connective tissue. Genes involved in the regulation of tissue remodeling are therefore candidate genes. We analyzed TIMP1 and TIMP2 coding sequences in 12 individuals with abdominal aortic aneurysms (AAA), one individual with AAA and intracranial aneurysms (IA), four individuals with IA and two clinically unaffected individuals. We identified two nucleotide variants in both the TIMP1 and the TIMP2 coding sequences. All differences occurred in the third base positions of codons and were neutral polymorphisms. A significant difference was observed in the frequency of TIMP2 nt 573 polymorphism between 168 alleles from AAA patients and 102 control alleles. [less ▲]Detailed reference viewed: 16 (1 ULg)