Hydra metalloproteinase 1: a secreted astacin metalloproteinase whose apical axis expression is differentially regulated during head regeneration.

The newly emerging astacin metalloproteinase family comprises multiple members with diverse functions. Most recently, the development-related functions have been attributed to both (1) proteolytic cleavage and subsequent release of active TGF-beta-like growth factors from latent inhibitory complexes and (2) modification of extracellular matrix (ECM) assembly and composition. We previously identified and purified hydra metalloproteinase 1 (HMP-1), a developmentally important astacin proteinase that functions in head regeneration and transdifferentiation of tentacle battery cells (L. Yan et al., 1995, Development 121, 1591-1602). In the present study, further cloning revealed that HMP-1 is produced as a secreted zymogen with a conserved hydrophobic signal sequence and a putative propeptide. The processed HMP-1 is composed of a characteristic astacin proteinase domain and a unique Cys-rich C-terminus. With this simple domain structure, HMP-1 represents an ancestral astacin proteinase. Consistent with its role in head regeneration, HMP-1 mRNA is expressed at highest levels by endodermal cells at the apical pole of the body column just inferior to the base of tentacles, the region of active cell differentiation or transdifferentiation. A modified immunocytochemical procedure demonstrated that HMP-1 protein can be localized not only to ECM of tentacles as we previously reported, but also to endodermal cells of the body column in a pattern similar to its mRNA distribution. The localization of HMP-1 protein in tentacles was confirmed using an enzymatic approach. A translocation of HMP-1 protein from cells in the body column to the extracellular milieu in tentacles further suggests that HMP-1 is a secreted protein. HMP-1 expression undergoes extensive regulation at the transcriptional level both temporally and spatially during head regeneration. The involvement of HMP-1 in this morphogenetic process is further supported by the blockage of head regeneration with localized antisense treatment. Taken together, these results suggest that HMP-1 is a secreted astacin metalloproteinase that has an important role in regulating hydra head morphogenesis potentially through its differential expression along the body axis.

[1]  C. Davis,et al.  The many faces of epidermal growth factor repeats. , 1990, The New biologist.

[2]  J. Palis,et al.  Early mRNAs, spatially restricted along the animal-vegetal axis of sea urchin embryos, include one encoding a protein related to tolloid and BMP-1. , 1992, Development.

[3]  E. Rossomando,et al.  Morphogenesis : an analysis of the development of biological form , 1992 .

[4]  H. Kung,et al.  Cloning and expression of cDNA encoding Xenopus laevis bone morphogenetic protein-1 during early embryonic development. , 1993, Gene.

[5]  J. Spring,et al.  A toxin homology domain in an astacin-like metalloproteinase of the jellyfish Podocoryne carnea with a dual role in digestion and development , 1998, Development Genes and Evolution.

[6]  S. Yasumasu,et al.  Isolation of cDNAs for LCE and HCE, two constituent proteases of the hatching enzyme of Oryzias latipes, and concurrent expression of their mRNAs during development. , 1992, Developmental biology.

[7]  P. S. St. John,et al.  Cloning and biological function of laminin in Hydra vulgaris. , 1994, Developmental biology.

[8]  M. Sarras.  BMP-1 and the astacin family of metalloproteinases: a potential link between the extracellular matrix, growth factors and pattern formation. , 1996, BioEssays : news and reviews in molecular, cellular and developmental biology.

[9]  K. Miyazono,et al.  Latent Transforming Growth Factor-/ l Associates to Fibroblast Extracellular Matrix via Latent ' II 3 FB Binding Protein , 2002 .

[10]  D. Greenspan,et al.  Characterization of a novel gene product (mammalian tolloid-like) with high sequence similarity to mammalian tolloid/bone morphogenetic protein-1. , 1996, Genomics.

[11]  W. Gelbart,et al.  An activity gradient of decapentaplegic is necessary for the specification of dorsal pattern elements in the Drosophila embryo. , 1993, Development.

[12]  Ken W. Y. Cho,et al.  Production of a DPP Activity Gradient in the Early Drosophila Embryo through the Opposing Actions of the SOG and TLD Proteins , 1997, Cell.

[13]  M. Sarras,et al.  Response to insulin and the expression pattern of a gene encoding an insulin receptor homologue suggest a role for an insulin-like molecule in regulating growth and patterning in Hydra , 1996, Development Genes and Evolution.

[14]  L. Stateva,et al.  The Astacins: structure and function of a new protein family , 1997 .

[15]  G. Lyons,et al.  Bone morphogenetic protein-1 and a mammalian tolloid homologue (mTld) are encoded by alternatively spliced transcripts which are differentially expressed in some tissues. , 1994, The Journal of biological chemistry.

[16]  N. Saitou,et al.  The neighbor-joining method: a new method for reconstructing phylogenetic trees. , 1987, Molecular biology and evolution.

[17]  P. S. St. John,et al.  Extracellular matrix (mesoglea) of Hydra vulgaris III. Formation and function during morphogenesis of hydra cell aggregates. , 1993, Developmental biology.

[18]  M. Sarras,et al.  Hydra cell aggregate development is blocked by selective fragments of fibronectin and type IV collagen. , 1994, Developmental biology.

[19]  L. Yan,et al.  A 25.7 x 10(3) M(r) hydra metalloproteinase (HMP1), a member of the astacin family, localizes to the extracellular matrix of Hydra vulgaris in a head-specific manner and has a developmental function. , 1995, Development.

[20]  W. Flanagan,et al.  Effects of oligonucleotide length, mismatches and mRNA levels on C-5 propyne-modified antisense potency. , 1996, Nucleic acids research.

[21]  S. Bryant,et al.  Pattern Formation: A Primer in Developmental Biology , 1984 .

[22]  V. Rosen,et al.  Novel regulators of bone formation: molecular clones and activities. , 1988 .

[23]  L. Yan,et al.  Identification and characterization of hydra metalloproteinase 2 (HMP2): a meprin-like astacin metalloproteinase that functions in foot morphogenesis. , 2000, Development.

[24]  K. Anderson,et al.  Localized enhancement and repression of the activity of the TGF-beta family member, decapentaplegic, is necessary for dorsal-ventral pattern formation in the Drosophila embryo. , 1992, Development.

[25]  R. Huber,et al.  Structure of astacin and implications for activation of astacins and zinc-ligation of collagenases , 1992, Nature.

[26]  D. Kingsley,et al.  What do BMPs do in mammals? Clues from the mouse short-ear mutation. , 1994, Trends in genetics : TIG.

[27]  M. Hentze,et al.  Determinants and regulation of cytoplasmic mRNA stability in eukaryotic cells. , 1991, Biochimica et biophysica acta.

[28]  Patrizia Ferretti,et al.  Cellular and Molecular Basis of Regeneration: From Invertebrates to Humans , 1998 .

[29]  M. O’Connor,et al.  Characterization of tolloid-related-1: a BMP-1-like product that is required during larval and pupal stages of Drosophila development. , 1994, Developmental biology.

[30]  D. Greenspan,et al.  Bone Morphogenetic Protein-1: The Type I Procollagen C-Proteinase , 1996, Science.

[31]  P. Bork,et al.  The CUB domain. A widespread module in developmentally regulated proteins. , 1993, Journal of molecular biology.

[32]  K. Titani,et al.  Amino acid sequence of a unique protease from the crayfish Astacus fluviatilis. , 1987, Biochemistry.

[33]  R. Beynon,et al.  The astacin family of metalloendopeptidases , 1991, The Journal of biological chemistry.

[34]  R. W. Padgett,et al.  Mutational analysis of the Drosophila tolloid gene, a human BMP-1 homolog. , 1994, Development.

[35]  T. Lepage,et al.  Spatial and temporal expression pattern during sea urchin embryogenesis of a gene coding for a protease homologous to the human protein BMP-1 and to the product of the Drosophila dorsal-ventral patterning gene tolloid. , 1992, Development.

[36]  F. Wardle,et al.  Regulation of BMP signaling by the BMP1/TLD-related metalloprotease, SpAN. , 1999, Developmental biology.

[37]  Embryonic expression of mouse bone morphogenetic protein-1 (BMP-1), which is related to the Drosophila dorsoventral gene tolloid and encodes a putative astacin metalloendopeptidase. , 1994, Developmental biology.

[38]  U. Strähle,et al.  Cleavage of the BMP-4 antagonist chordin by zebrafish tolloid. , 1997, Science.

[39]  K. Anderson,et al.  decapentaplegic acts as a morphogen to organize dorsal-ventral pattern in the Drosophila embryo , 1992, Cell.

[40]  W. Lennarz,et al.  Characterization of a homolog of human bone morphogenetic protein 1 in the embryo of the sea urchin, Strongylocentrotus purpuratus. , 1994, Development.

[41]  M. O’Connor,et al.  The Drosophila dorsal-ventral patterning gene tolloid is related to human bone morphogenetic protein 1 , 1991, Cell.

[42]  H. Bode,et al.  Evolutionary conservation of a cell fate specification gene: the Hydra achaete-scute homolog has proneural activity in Drosophila. , 1995, Development.

[43]  Davis Cg,et al.  The many faces of epidermal growth factor repeats. , 1990 .

[44]  M. Sarras,et al.  Extracellular matrix (mesoglea) of Hydra vulgaris. II. Influence of collagen and proteoglycan components on head regeneration. , 1991, Developmental biology.

[45]  B. Hogan,et al.  Growth factors in development: the role of TGF-beta related polypeptide signalling molecules in embryogenesis. , 1994, Development (Cambridge, England). Supplement.

[46]  D. Kingsley,et al.  The TGF-beta superfamily: new members, new receptors, and new genetic tests of function in different organisms. , 1994, Genes & development.

[47]  K. Anderson,et al.  Dorsal-ventral pattern formation in the Drosophila embryo: the role of zygotically active genes. , 1991, Current topics in developmental biology.

[48]  H. Bode,et al.  Transdifferentiation occurs continuously in adult hydra. , 1986, Current topics in developmental biology.

[49]  M. Sarras,et al.  Extracellular matrix (mesoglea) of Hydra vulgaris. I. Isolation and characterization. , 1991, Developmental biology.

[50]  G. Heijne.  A new method for predicting signal sequence cleavage sites. , 1986 .

[51]  D. Wilkinson.  In situ hybridization: a practical approach , 1998 .

[52]  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.

[53]  Leslie Dale,et al.  Cleavage of Chordin by Xolloid Metalloprotease Suggests a Role for Proteolytic Processing in the Regulation of Spemann Organizer Activity , 1997, Cell.

[54]  K. Miyazono,et al.  Latent transforming growth factor-beta 1 associates to fibroblast extracellular matrix via latent TGF-beta binding protein , 1994, The Journal of cell biology.

[55]  P. Chomczyński,et al.  Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. , 1987, Analytical biochemistry.

[56]  J. S. Bond,et al.  Role of the COOH-terminal Domains of Meprin A in Folding, Secretion, and Activity of the Metalloendopeptidase* , 1998, The Journal of Biological Chemistry.

[57]  M. Bernfield.  The basal lamina in epithelial-mesenchymal morphogenetic interactions. , 1977, Upsala journal of medical sciences.

[58]  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.

[59]  R. Harland,et al.  In situ hybridization: an improved whole-mount method for Xenopus embryos. , 1991, Methods in cell biology.