Cleavage of the BMP-4 Antagonist Chordin by Zebrafish Tolloid
10.1126/science.278.5345.1937
Crossref journal-article
American Association for the Advancement of Science (AAAS)
Science (221)
Abstract

Dorsoventral patterning of vertebrate and Drosophila embryos requires bone morphogenetic proteins (BMPs) and antagonists of BMP activity. The Drosophila gene tolloid encodes a metalloprotease similar to BMP-1 that interacts genetically with decapentaplegic , the Drosophila homolog of vertebrate BMP-2/4. Zebrafish embryos overexpressing a zebrafish homolog of tolloid were shown to resemble loss-of-function mutations in chordino , the zebrafish homolog of the Xenopus BMP-4 antagonist Chordin. Furthermore, Chordin was degraded by COS cells expressing Tolloid. These data suggest that Tolloid antagonizes Chordin activity by proteolytically cleaving Chordin. A conserved function for zebrafish and Drosophila Tolloid during embryogenesis is proposed.

Bibliography

Blader, P., Rastegar, S., Fischer, N., & Strähle, U. (1997). Cleavage of the BMP-4 Antagonist Chordin by Zebrafish Tolloid. Science, 278(5345), 1937–1940.

Authors 4
  1. Patrick Blader (first)
  2. Sepand Rastegar (additional)
  3. Nadine Fischer (additional)
  4. Uwe Strähle (additional)
References 47 Referenced 164
  1. J. M. W. Slack From Egg to Embryo (Cambridge Univ. Press Cambridge UK 1991); (10.1017/CBO9780511525322)
  2. Sive H. L., Genes Dev. 7, 1 (1993); (10.1101/gad.7.1.1) / Genes Dev. by Sive H. L. (1993)
  3. 10.1126/science.7939714
  4. Dale L., Howes G., Price B. M. L., Smith J. C., Development 115, 573 (1992). (10.1242/dev.115.2.573) / Development by Dale L. (1992)
  5. Hammerschmidt M., et al., ibid. 123, 95 (1996). / ibid. by Hammerschmidt M. (1996)
  6. 10.1016/S0925-4773(97)00659-X
  7. Hammerschmidt M., Serbedzija G. N., McMahon A. P., Genes Dev. 10, 2452 (1996). (10.1101/gad.10.19.2452) / Genes Dev. by Hammerschmidt M. (1996)
  8. Graff J. M., Thies R. S., Song J. J., Celeste A. J., Melton D. A., Cell 79, 169 (1994). (10.1016/0092-8674(94)90409-X) / Cell by Graff J. M. (1994)
  9. Y. Sasai et al. ibid. p. 779.
  10. Smith W. C., Harland R. M., ibid. 70, 829 (1992). / ibid. by Smith W. C. (1992)
  11. S. Piccolo Y. Sasai B. Lu E. M. De Robertis ibid. 86 589 (1996). (10.1016/S0092-8674(00)80132-4)
  12. L. B. Zimmerman J. M. De Jesus-Escobar R. M. Harland ibid. p. 599.
  13. Schulte-Merker S., Lee K. J., McMahon A. P., Hammerschmidt M., Nature 387, 862 (1997). (10.1038/43092) / Nature by Schulte-Merker S. (1997)
  14. Ferguson E. L., Anderson K. V., Development 114, 583 (1992). (10.1242/dev.114.3.583) / Development by Ferguson E. L. (1992)
  15. Padgett R. W., St. Johnson R. D., Gelbart W. M., Nature 325, 81 (1987); (10.1038/325081a0) / Nature by Padgett R. W. (1987)
  16. 10.1073/pnas.90.7.2905
  17. Holley S. A., et al., Nature 376, 249 (1995); (10.1038/376249a0) / Nature by Holley S. A. (1995)
  18. François V., Bier E., Cell 80, 19 (1995). (10.1016/0092-8674(95)90446-8) / Cell by François V. (1995)
  19. G. Jurgens E. Wieschaus C. Nüsslein-Volhard H. Kluding Wilhelm Roux's Arch. Dev. Biol. 193 283 (1984); C. Nüsslein-Volhard E. Wieschaus H. Kluding ibid. p. 267; B. T. Wakimoto (10.1007/BF00848156)
  20. Turner R. F., Kaufman T. C., Dev. Biol. 102, 147 (1984); (10.1016/0012-1606(84)90182-9) / Dev. Biol. by Turner R. F. (1984)
  21. ; E. Wieschaus C. Nüsslein-Volhard G. Jurgens Wilhelm Roux's Arch. Dev. Biol. 193 296 (1984); (10.1007/BF00848158)
  22. Zusman S. B., Wieschaus E., Dev. Biol. 111, 359 (1985). (10.1016/0012-1606(85)90490-7) / Dev. Biol. by Zusman S. B. (1985)
  23. M. J. Shimell E. L. Ferguson S. R. Childs M. B. O'Connor Cell 67 469 (1991). (10.1016/0092-8674(91)90522-Z)
  24. Sarras M. P., BioEssays 18, 439 (1996). (10.1002/bies.950180604) / BioEssays by Sarras M. P. (1996)
  25. 10.1126/science.3201241
  26. Li S.-W., et al., Proc. Natl. Acad. Sci. U.S.A. 93, 5127 (1996); (10.1073/pnas.93.10.5127) / Proc. Natl. Acad. Sci. U.S.A. by Li S.-W. (1996)
  27. 10.1126/science.271.5247.360
  28. Takahara K., Lyons G. E., Greenspan D. S., J. Biol. Chem. 269, 32572 (1994); (10.1016/S0021-9258(18)31672-7) / J. Biol. Chem. by Takahara K. (1994)
  29. Fukagawa M., Suzuki N., Hogan B. L. M., Jones C. M., Dev. Biol. 163, 175 (1994). (10.1006/dbio.1994.1133) / Dev. Biol. by Fukagawa M. (1994)
  30. A partial cDNA fragment encoding part of the metalloprotease domain of a zebrafish Tolloid homolog was isolated by reverse transcriptase–polymerase chain reaction (RT-PCR) with RNA from 24-hour embryos as template and the following oligonucleotide primers: GCTACCAGAATTCGCIATGIGICA(C/T)TGGGA and AGTACGGGATCCITTICIIGC(A/G)TA(A/G)TGCAT. The PCR fragment was used as a probe against a gastrula stage cDNA library and several positive phage were purified and their inserts liberated by in vitro excision. The plasmid with the largest cDNA insert (GenBank accession number ) was sequenced on both strands.
  31. In situ hybridization with digoxygenin-incorporated antisense RNA probes was done as decribed [
  32. Oxtoby E., Jowett T., Nucleic Acids Res. 21, 1087 (1993); (10.1093/nar/21.5.1087) / Nucleic Acids Res. by Oxtoby E. (1993)
  33. ]. Fish embryo raising and staging were as described [M. Westerfield The Zebrafish Book (Univ. of Oregon Press Eugene 1995)].
  34. P. Blader S. Rastegar N. Fischer U. Strähle data not shown.
  35. Wild-type and mutated zebrafish tolloid injection constructs were generated by PCR in two steps with the high-fidelity Pfu polymerase (Stratagene) to avoid the introduction of amplification artifacts. DNA encoding a region from the initiation methionine to the end of the metalloprotease domain was amplified and cloned into pCS2+ to generate pCS2:Metallo; the corresponding mutated construct pCS2:Metallo Mut was assembled from two PCR fragments in which internal oligonucleotide primers were designed to introduce amino acid substitutions in the resultant zinc-binding domain. Subsequently a PCR fragment encoding the COOH-terminal portion of zebrafish Tolloid was subcloned into pCS2:Metallo or pCS2:Metallo Mut . The two resultant full-length constructs pCS2:Tolloid and pCS2:Tolloid Mut were linearized with Not I and transcribed with the Message Machine Kit (Ambion). For transfection into COS cells constructs were tagged at the COOH-terminus with the Myc epitope. The integrity of synthetic RNAs was assessed on agarose gels and the RNAs were diluted in water to their respective final injection concentrations immediately before injection. Synthetic mRNAs were injected into the yolk of one- to two-cell-stage embryos with a gas-driven microinjector (Eppendorf).
  36. Joly J. S., Joly C., Schultemerker S., Boulekbache H., Condamine H., Development 119, 1261 (1993). (10.1242/dev.119.4.1261) / Development by Joly J. S. (1993)
  37. Detrich H. W., et al., Proc. Natl. Acad. Sci. U.S.A. 92, 10713 (1995). (10.1073/pnas.92.23.10713) / Proc. Natl. Acad. Sci. U.S.A. by Detrich H. W. (1995)
  38. Krauss S., Johansen T., Korzh V., Fjose A., Development 113, 1193 (1991); (10.1242/dev.113.4.1193) / Development by Krauss S. (1991)
  39. Püschel A. W., Westerfield M., Dressler G. R., Mech. Dev. 38, 197 (1992). (10.1016/0925-4773(92)90053-M) / Mech. Dev. by Püschel A. W. (1992)
  40. Finelli A. L., Bossie C. A., Xie T., Padgett R. W., Development 120, 861 (1994). (10.1242/dev.120.4.861) / Development by Finelli A. L. (1994)
  41. Mullins M. C., et al., ibid. 123, 81 (1996). / ibid. by Mullins M. C. (1996)
  42. Epitope-tagged Chordin (Chd-MYC) was produced from a baculovirus construct with the use of High-Five cells (Invitrogen) (9) and Myc immunoreactivity was subsequently monitored by protein immunoblotting with monoclonal antibody to Myc (9E10) [
  43. Evans G. L., Lewis G. K., Ramsey G., Bishop J. M., Mol. Cell. Biol. 5, 3610 (1985); / Mol. Cell. Biol. by Evans G. L. (1985)
  44. ]. Purified retinoid X receptor α–ligand binding domain (RXRα-LBD) was used as a control for protease specificity [W. Bourguet et al. Protein Express. Purif. 6 604 (1995)]. COS cells were transfected with various DNA constructs with DEAE dextran as described [
  45. Cullen B. R., Methods Enzymol. 152, 684 (1987); (10.1016/0076-6879(87)52074-2) / Methods Enzymol. by Cullen B. R. (1987)
  46. ]. Expression of the transfected constructs was monitored in parallel by immunohistocytochemistry. The proteolytic potential of the transfected cells was assayed after 48 hours in culture by removing half of the growth media from the COS cells and replacing it with an equal volume of prewarmed baculovirus extract supplemented with RXRα-LBD. Samples of the resulting media were removed at various times after addition of the substrates and were assayed for immunoreactivity by protein immunoblotting. Proteins from the transfected constructs were expressed at a level insufficient for detection by protein immunoblotting of the COS supernatants.
  47. We are indebted to H.-F. Kung for providing the DN-BR expression construct R. Harland for providing the Xenopus noggin expression construct E. M. De Robertis for providing the Xenopus Chordin expression plasmid and Chd-MYC–expressing baculovirus P. Egea for providing the purified RXRα-LBD and anti-RXRα-LBD and J. S. Joly S. Krauss and L. Zon for probes. We thank the staff of the fish and cell culture facilities the photographers the oligosynthesis and sequencing groups of the IGBMC and N. Foulkes for technical assistance and critical reading of the manuscript. P.B. was supported by a Training and Mobility for Researchers (TMR) fellowship from the European Community. U.S. was the recipient of a fellowship from the Deutsche Forschungsgemeinschaft and the Centre International des Etudiants et Stagaires (CIES). Supported by the Institut National de la Santé et de la Recherche Médicale the Centre National de la Recherche Scientifique the Centre Hospitalier Universitaire Régional the Association pour la Recherche sur le Cancer (ARC) the Groupement de Recherche et d'Etudes sur les Génomes (GREG) the Association Français coutre les Myopathies (AFM) and La Ligue contre le Cancer.
Dates
Type When
Created 23 years, 1 month ago (July 27, 2002, 5:37 a.m.)
Deposited 1 year, 7 months ago (Jan. 12, 2024, 11:44 p.m.)
Indexed 1 month, 1 week ago (July 24, 2025, 8:05 a.m.)
Issued 27 years, 8 months ago (Dec. 12, 1997)
Published 27 years, 8 months ago (Dec. 12, 1997)
Published Print 27 years, 8 months ago (Dec. 12, 1997)
Funders 0

None

@article{Blader_1997, title={Cleavage of the BMP-4 Antagonist Chordin by Zebrafish Tolloid}, volume={278}, ISSN={1095-9203}, url={http://dx.doi.org/10.1126/science.278.5345.1937}, DOI={10.1126/science.278.5345.1937}, number={5345}, journal={Science}, publisher={American Association for the Advancement of Science (AAAS)}, author={Blader, Patrick and Rastegar, Sepand and Fischer, Nadine and Strähle, Uwe}, year={1997}, month=dec, pages={1937–1940} }