Skip to main content
PMC home page
The EMBO Journal logoLink to The EMBO Journal
. 1990 Sep;9(9):3003–3012. doi: 10.1002/j.1460-2075.1990.tb07493.x

Early expression of a collagenase-like hatching enzyme gene in the sea urchin embryo.

T Lepage 1, C Gache 1
PMCID: PMC552018  PMID: 2167841

Abstract

The hatching enzyme is a developmentally regulated protease secreted at the blastula stage by the sea urchin embryo to digest its protective envelope. A nearly full-length cDNA clone (HE6) encoding the entire sequence of the hatching enzyme was isolated from a prehatching blastula lambda gt11 cDNA library. The 1761 bp open reading frame codes for a preprohatching enzyme with an 18 amino acid signal sequence, a 148 amino acid activation peptide and a 421 amino acid mature enzyme which has homologies with the mammalian collagenases. Transcripts of the hatching enzyme gene are not detected in the unfertilized egg, they accumulate during the cleavage stages and disappear at hatching. This transient expression results from a transcriptional control. Thus the hatching enzyme mRNA is not a maternal gene product but a transcript synthesized at a very early stage from the zygotic genome.

Full text

PDF
3006

Images in this article

3006Fig. 3.
on p.3006
3008Fig. 6.
on p.3008
3008Fig. 7.
on p.3008

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Aebersold R. H., Teplow D. B., Hood L. E., Kent S. B. Electroblotting onto activated glass. High efficiency preparation of proteins from analytical sodium dodecyl sulfate-polyacrylamide gels for direct sequence analysis. J Biol Chem. 1986 Mar 25;261(9):4229–4238. [PubMed] [Google Scholar]
  2. Alexander C. M., Werb Z. Proteinases and extracellular matrix remodeling. Curr Opin Cell Biol. 1989 Oct;1(5):974–982. doi: 10.1016/0955-0674(89)90068-9. [DOI] [PubMed] [Google Scholar]
  3. Angel P., Imagawa M., Chiu R., Stein B., Imbra R. J., Rahmsdorf H. J., Jonat C., Herrlich P., Karin M. Phorbol ester-inducible genes contain a common cis element recognized by a TPA-modulated trans-acting factor. Cell. 1987 Jun 19;49(6):729–739. doi: 10.1016/0092-8674(87)90611-8. [DOI] [PubMed] [Google Scholar]
  4. Angerer L. M., Chambers S. A., Yang Q., Venkatesan M., Angerer R. C., Simpson R. T. Expression of a collagen gene in mesenchyme lineages of the Strongylocentrotus purpuratus embryo. Genes Dev. 1988 Feb;2(2):239–246. doi: 10.1101/gad.2.2.239. [DOI] [PubMed] [Google Scholar]
  5. Aviv H., Leder P. Purification of biologically active globin messenger RNA by chromatography on oligothymidylic acid-cellulose. Proc Natl Acad Sci U S A. 1972 Jun;69(6):1408–1412. doi: 10.1073/pnas.69.6.1408. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Barrett D., Angelo G. M. Maternal characteristics of hatching enzymes in hybrid sea urchin embryos. Exp Cell Res. 1969 Oct;57(2):159–166. doi: 10.1016/0014-4827(69)90137-2. [DOI] [PubMed] [Google Scholar]
  7. Barrett D., Edwards B. F. Hatching enzyme of the sea urchin Strongylocentrotus purpuratus. Methods Enzymol. 1976;45:354–373. doi: 10.1016/s0076-6879(76)45032-2. [DOI] [PubMed] [Google Scholar]
  8. Benson S., Sucov H., Stephens L., Davidson E., Wilt F. A lineage-specific gene encoding a major matrix protein of the sea urchin embryo spicule. I. Authentication of the cloned gene and its developmental expression. Dev Biol. 1987 Apr;120(2):499–506. doi: 10.1016/0012-1606(87)90253-3. [DOI] [PubMed] [Google Scholar]
  9. Brenner C. A., Adler R. R., Rappolee D. A., Pedersen R. A., Werb Z. Genes for extracellular-matrix-degrading metalloproteinases and their inhibitor, TIMP, are expressed during early mammalian development. Genes Dev. 1989 Jun;3(6):848–859. doi: 10.1101/gad.3.6.848. [DOI] [PubMed] [Google Scholar]
  10. Britten R. J., Cetta A., Davidson E. H. The single-copy DNA sequence polymorphism of the sea urchin Strongylocentrotus purpuratus. Cell. 1978 Dec;15(4):1175–1186. doi: 10.1016/0092-8674(78)90044-2. [DOI] [PubMed] [Google Scholar]
  11. Bruskin A. M., Tyner A. L., Wells D. E., Showman R. M., Klein W. H. Accumulation in embryogenesis of five mRNAs enriched in the ectoderm of the sea urchin pluteus. Dev Biol. 1981 Oct 30;87(2):308–318. doi: 10.1016/0012-1606(81)90154-8. [DOI] [PubMed] [Google Scholar]
  12. Cavener D. R. Comparison of the consensus sequence flanking translational start sites in Drosophila and vertebrates. Nucleic Acids Res. 1987 Feb 25;15(4):1353–1361. doi: 10.1093/nar/15.4.1353. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Chiu R., Boyle W. J., Meek J., Smeal T., Hunter T., Karin M. The c-Fos protein interacts with c-Jun/AP-1 to stimulate transcription of AP-1 responsive genes. Cell. 1988 Aug 12;54(4):541–552. doi: 10.1016/0092-8674(88)90076-1. [DOI] [PubMed] [Google Scholar]
  14. Chou P. Y., Fasman G. D. Empirical predictions of protein conformation. Annu Rev Biochem. 1978;47:251–276. doi: 10.1146/annurev.bi.47.070178.001343. [DOI] [PubMed] [Google Scholar]
  15. Collier I. E., Wilhelm S. M., Eisen A. Z., Marmer B. L., Grant G. A., Seltzer J. L., Kronberger A., He C. S., Bauer E. A., Goldberg G. I. H-ras oncogene-transformed human bronchial epithelial cells (TBE-1) secrete a single metalloprotease capable of degrading basement membrane collagen. J Biol Chem. 1988 May 15;263(14):6579–6587. [PubMed] [Google Scholar]
  16. Costanzi C., Gillespie D. Fast blots: immobilization of DNA and RNA from cells. Methods Enzymol. 1987;152:582–587. doi: 10.1016/0076-6879(87)52065-1. [DOI] [PubMed] [Google Scholar]
  17. Devault A., Lazure C., Nault C., Le Moual H., Seidah N. G., Chrétien M., Kahn P., Powell J., Mallet J., Beaumont A. Amino acid sequence of rabbit kidney neutral endopeptidase 24.11 (enkephalinase) deduced from a complementary DNA. EMBO J. 1987 May;6(5):1317–1322. doi: 10.1002/j.1460-2075.1987.tb02370.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Dolecki G. J., Wannakrairoj S., Lum R., Wang G., Riley H. D., Carlos R., Wang A., Humphreys T. Stage-specific expression of a homeo box-containing gene in the non-segmented sea urchin embryo. EMBO J. 1986 May;5(5):925–930. doi: 10.1002/j.1460-2075.1986.tb04305.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Edelman A. M., Blumenthal D. K., Krebs E. G. Protein serine/threonine kinases. Annu Rev Biochem. 1987;56:567–613. doi: 10.1146/annurev.bi.56.070187.003031. [DOI] [PubMed] [Google Scholar]
  20. Feinberg A. P., Vogelstein B. A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity. Anal Biochem. 1983 Jul 1;132(1):6–13. doi: 10.1016/0003-2697(83)90418-9. [DOI] [PubMed] [Google Scholar]
  21. GROSS P. R., COUSINEAU G. H. Effects of actinomycin D on macromolecule synthesis and early development in sea urchin eggs. Biochem Biophys Res Commun. 1963 Feb 18;10:321–326. doi: 10.1016/0006-291x(63)90532-1. [DOI] [PubMed] [Google Scholar]
  22. Garfinkel M. D., Pruitt R. E., Meyerowitz E. M. DNA sequences, gene regulation and modular protein evolution in the Drosophila 68C glue gene cluster. J Mol Biol. 1983 Aug 25;168(4):765–789. doi: 10.1016/s0022-2836(83)80074-6. [DOI] [PubMed] [Google Scholar]
  23. Garnier J., Osguthorpe D. J., Robson B. Analysis of the accuracy and implications of simple methods for predicting the secondary structure of globular proteins. J Mol Biol. 1978 Mar 25;120(1):97–120. doi: 10.1016/0022-2836(78)90297-8. [DOI] [PubMed] [Google Scholar]
  24. Goldberg G. I., Wilhelm S. M., Kronberger A., Bauer E. A., Grant G. A., Eisen A. Z. Human fibroblast collagenase. Complete primary structure and homology to an oncogene transformation-induced rat protein. J Biol Chem. 1986 May 15;261(14):6600–6605. [PubMed] [Google Scholar]
  25. Grant S. R., Farach M. C., Decker G. L., Woodward H. D., Farach H. A., Jr, Lennarz W. J. Developmental expression of cell-surface (glyco)proteins involved in gastrulation and spicule formation in sea urchin embryos. Cold Spring Harb Symp Quant Biol. 1985;50:91–98. doi: 10.1101/sqb.1985.050.01.013. [DOI] [PubMed] [Google Scholar]
  26. Gubler U., Hoffman B. J. A simple and very efficient method for generating cDNA libraries. Gene. 1983 Nov;25(2-3):263–269. doi: 10.1016/0378-1119(83)90230-5. [DOI] [PubMed] [Google Scholar]
  27. Harkey M. A., Whiteley H. R., Whiteley A. H. Coordinate accumulation of five transcripts in the primary mesenchyme during skeletogenesis in the sea urchin embryo. Dev Biol. 1988 Feb;125(2):381–395. doi: 10.1016/0012-1606(88)90219-9. [DOI] [PubMed] [Google Scholar]
  28. Hattori K., Angel P., Le Beau M. M., Karin M. Structure and chromosomal localization of the functional intronless human JUN protooncogene. Proc Natl Acad Sci U S A. 1988 Dec;85(23):9148–9152. doi: 10.1073/pnas.85.23.9148. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Henikoff S. Unidirectional digestion with exonuclease III creates targeted breakpoints for DNA sequencing. Gene. 1984 Jun;28(3):351–359. doi: 10.1016/0378-1119(84)90153-7. [DOI] [PubMed] [Google Scholar]
  30. Jongeneel C. V., Bouvier J., Bairoch A. A unique signature identifies a family of zinc-dependent metallopeptidases. FEBS Lett. 1989 Jan 2;242(2):211–214. doi: 10.1016/0014-5793(89)80471-5. [DOI] [PubMed] [Google Scholar]
  31. Kester W. R., Matthews B. W. Comparison of the structures of carboxypeptidase A and thermolysin. J Biol Chem. 1977 Nov 10;252(21):7704–7710. [PubMed] [Google Scholar]
  32. Kozak M. An analysis of 5'-noncoding sequences from 699 vertebrate messenger RNAs. Nucleic Acids Res. 1987 Oct 26;15(20):8125–8148. doi: 10.1093/nar/15.20.8125. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Kozak M. The scanning model for translation: an update. J Cell Biol. 1989 Feb;108(2):229–241. doi: 10.1083/jcb.108.2.229. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Lawler J., Hynes R. O. The structure of human thrombospondin, an adhesive glycoprotein with multiple calcium-binding sites and homologies with several different proteins. J Cell Biol. 1986 Nov;103(5):1635–1648. doi: 10.1083/jcb.103.5.1635. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Lee J. J., Calzone F. J., Britten R. J., Angerer R. C., Davidson E. H. Activation of sea urchin actin genes during embryogenesis. Measurement of transcript accumulation from five different genes in Strongylocentrotus purpuratus. J Mol Biol. 1986 Mar 20;188(2):173–183. doi: 10.1016/0022-2836(86)90302-5. [DOI] [PubMed] [Google Scholar]
  36. Lepage T., Gache C. Purification and characterization of the sea urchin embryo hatching enzyme. J Biol Chem. 1989 Mar 25;264(9):4787–4793. [PubMed] [Google Scholar]
  37. Lipman D. J., Pearson W. R. Rapid and sensitive protein similarity searches. Science. 1985 Mar 22;227(4693):1435–1441. doi: 10.1126/science.2983426. [DOI] [PubMed] [Google Scholar]
  38. Marck C. 'DNA Strider': a 'C' program for the fast analysis of DNA and protein sequences on the Apple Macintosh family of computers. Nucleic Acids Res. 1988 Mar 11;16(5):1829–1836. doi: 10.1093/nar/16.5.1829. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Matrisian L. M., Leroy P., Ruhlmann C., Gesnel M. C., Breathnach R. Isolation of the oncogene and epidermal growth factor-induced transin gene: complex control in rat fibroblasts. Mol Cell Biol. 1986 May;6(5):1679–1686. doi: 10.1128/mcb.6.5.1679. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Mierendorf R. C., Percy C., Young R. A. Gene isolation by screening lambda gt11 libraries with antibodies. Methods Enzymol. 1987;152:458–469. doi: 10.1016/0076-6879(87)52054-7. [DOI] [PubMed] [Google Scholar]
  41. Mierendorf R. C., Pfeffer D. Direct sequencing of denatured plasmid DNA. Methods Enzymol. 1987;152:556–562. doi: 10.1016/0076-6879(87)52061-4. [DOI] [PubMed] [Google Scholar]
  42. Morris G. F., Marzluff W. F. A factor in sea urchin eggs inhibits transcription in isolated nuclei by sea urchin RNA polymerase III. Biochemistry. 1983 Feb 1;22(3):645–653. doi: 10.1021/bi00272a019. [DOI] [PubMed] [Google Scholar]
  43. Murray A. W., Kirschner M. W. Cyclin synthesis drives the early embryonic cell cycle. Nature. 1989 May 25;339(6222):275–280. doi: 10.1038/339275a0. [DOI] [PubMed] [Google Scholar]
  44. Nevins J. R. Isolation and analysis of nuclear RNA. Methods Enzymol. 1987;152:234–241. doi: 10.1016/0076-6879(87)52025-0. [DOI] [PubMed] [Google Scholar]
  45. Nocente-McGrath C., Brenner C. A., Ernst S. G. Endo16, a lineage-specific protein of the sea urchin embryo, is first expressed just prior to gastrulation. Dev Biol. 1989 Nov;136(1):264–272. doi: 10.1016/0012-1606(89)90147-4. [DOI] [PubMed] [Google Scholar]
  46. Olmsted J. B. Affinity purification of antibodies from diazotized paper blots of heterogeneous protein samples. J Biol Chem. 1981 Dec 10;256(23):11955–11957. [PubMed] [Google Scholar]
  47. Pearson W. R., Lipman D. J. Improved tools for biological sequence comparison. Proc Natl Acad Sci U S A. 1988 Apr;85(8):2444–2448. doi: 10.1073/pnas.85.8.2444. [DOI] [PMC free article] [PubMed] [Google Scholar]
  48. Ringwald M., Schuh R., Vestweber D., Eistetter H., Lottspeich F., Engel J., Dölz R., Jähnig F., Epplen J., Mayer S. The structure of cell adhesion molecule uvomorulin. Insights into the molecular mechanism of Ca2+-dependent cell adhesion. EMBO J. 1987 Dec 1;6(12):3647–3653. doi: 10.1002/j.1460-2075.1987.tb02697.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  49. Ruoslahti E. Fibronectin and its receptors. Annu Rev Biochem. 1988;57:375–413. doi: 10.1146/annurev.bi.57.070188.002111. [DOI] [PubMed] [Google Scholar]
  50. Sanchez-Lopez R., Nicholson R., Gesnel M. C., Matrisian L. M., Breathnach R. Structure-function relationships in the collagenase family member transin. J Biol Chem. 1988 Aug 25;263(24):11892–11899. [PubMed] [Google Scholar]
  51. Sanger F., Nicklen S., Coulson A. R. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. doi: 10.1073/pnas.74.12.5463. [DOI] [PMC free article] [PubMed] [Google Scholar]
  52. Schönthal A., Herrlich P., Rahmsdorf H. J., Ponta H. Requirement for fos gene expression in the transcriptional activation of collagenase by other oncogenes and phorbol esters. Cell. 1988 Jul 29;54(3):325–334. doi: 10.1016/0092-8674(88)90195-x. [DOI] [PubMed] [Google Scholar]
  53. Shaw G., Kamen R. A conserved AU sequence from the 3' untranslated region of GM-CSF mRNA mediates selective mRNA degradation. Cell. 1986 Aug 29;46(5):659–667. doi: 10.1016/0092-8674(86)90341-7. [DOI] [PubMed] [Google Scholar]
  54. Showman R. M., Foerder C. A. Removal of the fertilization membrane of sea urchin embryos employing aminotriazole. Exp Cell Res. 1979 May;120(2):253–255. doi: 10.1016/0014-4827(79)90385-9. [DOI] [PubMed] [Google Scholar]
  55. Soeiro R., Darnell J. E. Competition hybridization by "pre-saturation" of HeLa cell DNA. J Mol Biol. 1969 Sep 28;44(3):551–562. doi: 10.1016/0022-2836(69)90379-9. [DOI] [PubMed] [Google Scholar]
  56. Spinelli G., Gianguzza F., Casano C., Acierno P., Burckhardt J. Evidences of two different sets of histone genes active during embryogenesis of the sea urchin Paracentrotus lividus. Nucleic Acids Res. 1979 Feb;6(2):545–560. doi: 10.1093/nar/6.2.545. [DOI] [PMC free article] [PubMed] [Google Scholar]
  57. Stricklin G. P., Jeffrey J. J., Roswit W. T., Eisen A. Z. Human skin fibroblast procollagenase: mechanisms of activation by organomercurials and trypsin. Biochemistry. 1983 Jan 4;22(1):61–68. doi: 10.1021/bi00270a009. [DOI] [PubMed] [Google Scholar]
  58. Stuart D. I., Acharya K. R., Walker N. P., Smith S. G., Lewis M., Phillips D. C. Alpha-lactalbumin possesses a novel calcium binding loop. Nature. 1986 Nov 6;324(6092):84–87. doi: 10.1038/324084a0. [DOI] [PubMed] [Google Scholar]
  59. Tabor S., Richardson C. C. DNA sequence analysis with a modified bacteriophage T7 DNA polymerase. Proc Natl Acad Sci U S A. 1987 Jul;84(14):4767–4771. doi: 10.1073/pnas.84.14.4767. [DOI] [PMC free article] [PubMed] [Google Scholar]
  60. Takahashi N., Takahashi Y., Putnam F. W. Structure of human hemopexin: O-glycosyl and N-glycosyl sites and unusual clustering of tryptophan residues. Proc Natl Acad Sci U S A. 1984 Apr;81(7):2021–2025. doi: 10.1073/pnas.81.7.2021. [DOI] [PMC free article] [PubMed] [Google Scholar]
  61. Takeuchi K., Yokosawa H., Hoshi M. Purification and characterization of hatching enzyme of Strongylocentrotus intermedius. Eur J Biochem. 1979 Oct;100(1):257–265. doi: 10.1111/j.1432-1033.1979.tb02056.x. [DOI] [PubMed] [Google Scholar]
  62. Terman S. A., Gross P. R. Translation-level control of protein synthesis during early development. Biochem Biophys Res Commun. 1965 Dec 21;21(6):595–600. doi: 10.1016/0006-291x(65)90527-9. [DOI] [PubMed] [Google Scholar]
  63. Uzman J. A., Wilt F. H. The role of RNA polymerase initiation and elongation in control of total RNA and histone mRNA synthesis in sea urchin embryos. Dev Biol. 1984 Nov;106(1):174–180. doi: 10.1016/0012-1606(84)90073-3. [DOI] [PubMed] [Google Scholar]
  64. Wessel G. M., McClay D. R. Gastrulation in the sea urchin embryo requires the deposition of crosslinked collagen within the extracellular matrix. Dev Biol. 1987 May;121(1):149–165. doi: 10.1016/0012-1606(87)90148-5. [DOI] [PubMed] [Google Scholar]
  65. Whiteley H. R., Whiteley A. H. Changing populations of reiterated DNA transcripts during early echinoderm development. Curr Top Dev Biol. 1975;9:39–88. doi: 10.1016/s0070-2153(08)60026-5. [DOI] [PubMed] [Google Scholar]
  66. Whitham S. E., Murphy G., Angel P., Rahmsdorf H. J., Smith B. J., Lyons A., Harris T. J., Reynolds J. J., Herrlich P., Docherty A. J. Comparison of human stromelysin and collagenase by cloning and sequence analysis. Biochem J. 1986 Dec 15;240(3):913–916. doi: 10.1042/bj2400913. [DOI] [PMC free article] [PubMed] [Google Scholar]
  67. Yang Q., Angerer L. M., Angerer R. C. Structure and tissue-specific developmental expression of a sea urchin arylsulfatase gene. Dev Biol. 1989 Sep;135(1):53–65. doi: 10.1016/0012-1606(89)90157-7. [DOI] [PubMed] [Google Scholar]
  68. Yang Q., Angerer L. M., Angerer R. C. Unusual pattern of accumulation of mRNA encoding EGF-related protein in sea urchin embryos. Science. 1989 Nov 10;246(4931):806–808. doi: 10.1126/science.2814501. [DOI] [PubMed] [Google Scholar]
  69. Young R. A., Davis R. W. Yeast RNA polymerase II genes: isolation with antibody probes. Science. 1983 Nov 18;222(4625):778–782. doi: 10.1126/science.6356359. [DOI] [PubMed] [Google Scholar]
  70. von Heijne G. A new method for predicting signal sequence cleavage sites. Nucleic Acids Res. 1986 Jun 11;14(11):4683–4690. doi: 10.1093/nar/14.11.4683. [DOI] [PMC free article] [PubMed] [Google Scholar]
  71. von Heijne G. Signal sequences. The limits of variation. J Mol Biol. 1985 Jul 5;184(1):99–105. doi: 10.1016/0022-2836(85)90046-4. [DOI] [PubMed] [Google Scholar]

Articles from The EMBO Journal are provided here courtesy of Nature Publishing Group

RESOURCES