Skip to main content
SpringerLink
Log in

A signal peptide secretion screen in Fucus distichus embryos reveals expression of glucanase, EGF domain-containing, and LRR receptor kinase-like polypeptides during asymmetric cell growth

  • Original Article
  • Published:
Planta Aims and scope Submit manuscript

Abstract

Zygotes of the brown alga Fucus distichus (L.) Powell develop polarity prior to the first embryonic cell division and retain a pattern of asymmetric growth during early embryogenesis. In order to identify F. distichus polypeptides secreted during asymmetric cell growth, we used a functional assay in Saccharomyces cerevisiae to screen a cDNA library generated from asymmetrically growing Fucus embryos for sequences encoding polypeptides that function as signal peptides for secretion. We isolated and sequenced 222 plasmids containing Fucus cDNAs encoding signal peptide activity. The cDNA inserts from these plasmids were translated in silico into 244 potential polypeptide sequences, 169 of which are predicted to contain signal peptides. BlastP analysis of the Fucus sequences revealed similarity between many Fucus proteins and cell surface proteins that function in development in other eukaryotes, including epidermal growth factor (EGF)-like repeat-containing proteins, plant leucine-rich repeat (LRR)-receptor kinases, and algal β-1, 3-exoglucanase. However, most of the isolated Fucus polypeptides lack similarity to known proteins. The isolation of cDNAs encoding secreted Fucus proteins provides an important step toward characterizing cell surface proteins important for asymmetric organization and growth in fucoid embryos.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1a, b.
Fig. 2a, b.
Fig. 3a–c.

Similar content being viewed by others

Abbreviations

ECM:

extracellular matrix

EGF:

epidermal growth factor

ER:

endoplasmic reticulum

LRR:

leucine-rich repeat

SST:

signal sequence trap

WAK:

wall-associated kinase

References

  • Altschul SF, Gish W, Miller W, Myers EW, Lipman DJ (1990) Basic local alignment search tool. J Mol Biol. 215:403–410

  • An YQ, McDowell JM, Huang S, McKinney EC, Chambliss S, Meagher RB (1996) Strong, constitutive expression of the Arabidopsis ACT2/ACT8 actin subclass in vegetative tissues. Plant J 10:107–121

    CAS  PubMed  Google Scholar 

  • Appel RD, Bairoch A, Hochstrasser DF (1994) A new generation of information retrieval tools for biologists: the example of the ExPASy WWW server. Trends Biochem.Sci 19:258–260

    CAS  Google Scholar 

  • Apt KE, Clendennen SK, Powers DA, Grossman AR (1995) The gene family encoding the fucoxanthin chlorophyll proteins from the brown alga Macrocystis pyrifera. Mol Gen Genet 246:455–464

    CAS  PubMed  Google Scholar 

  • Ausubel FM, Brent R, Kingston RE, Moore DD, Seidman JG, Smith JA, Struhl KJ (eds) (1995) Current protocols in molecular biology. Wiley, New York, chpt 13

  • Belanger KD, Quatrano RS (2000a) Polarity: the role of localized secretion. Curr Opin Plant Biol 3:67–72

    Article  CAS  PubMed  Google Scholar 

  • Belanger KD, Quatrano RS (2000b) Membrane recycling occurs during asymmetric tip growth and cell plate formation in Fucus distichus zygotes. Protoplasma 212:24–37

    Google Scholar 

  • Bork P, Downing AK, Kieffer B, Campbell ID (1996) Structure and distribution of modules in extracellular proteins. Q Rev Biophys 29:119–167

    CAS  PubMed  Google Scholar 

  • Brawley SH, Quatrano RS (1979) Sulfation of fucoidin in Fucus embryos. 4. Autoradiographic investigations of fucoidin sulfation and secretion during differentiation and the effects of cytochalasin treatment. Dev Biol 73:193–205

    CAS  PubMed  Google Scholar 

  • Brownlee C, Bouget FY (1998) Polarity determination in Fucus: from zygote to multicellular embryo. Semin Cell Dev Biol 9:179–185

    Article  CAS  PubMed  Google Scholar 

  • Brownlee C, Bouget F-Y, Corellou F (2001) Choosing sides: establishment of polarity in zygotes of fucoid algae. Semin Cell Dev Biol 12:345–351

    Article  CAS  PubMed  Google Scholar 

  • Cheong YH, Kim CY, Chun HJ, Moon BC, Park HC, Kim JK, Lee SH, Han CD, Lee SY, Cho MJ (2000) Molecular cloning of a soybean class III beta-1,3-glucanase gene that is regulated both developmentally and in response to pathogen infection. Plant Sci 154:71–81

    Article  CAS  PubMed  Google Scholar 

  • Cove DJ (2000) The generation and modification of cell polarity. J Exp Bot 51:831–838

    Article  CAS  PubMed  Google Scholar 

  • Doe CQ, Bowerman B (2001) Asymmetric cell division: fly neuroblast meets worm zygote. Curr Opin Cell Biol 13:68–75

    Article  CAS  PubMed  Google Scholar 

  • Drgonova J, Drgon T, Tanaka K, Kollar R, Chen GC, Ford RA, Chan CS, Takai Y, Cabib E (1996) Rho1p, a yeast protein at the interface between cell polarization and morphogenesis. Science 272:277–279

    PubMed  Google Scholar 

  • Drubin DG, Nelson WJ (1996) Origins of cell polarity. Cell 84:335–344

    CAS  PubMed  Google Scholar 

  • Emanuelsson O, Nielsen H, von Heijne G (1999) ChloroP, a neural network-based method for predicting chloroplast transit peptides and their cleavage sites. Protein Sci 8:978–984

    PubMed  Google Scholar 

  • Gollan L, Sabanay H, Poliak S, Berglund EO, Ranscht B, Peles E (2002) Retention of a cell adhesion complex at the paranodal junction requires the cytoplasmic region of Caspr. J Cell Biol 157:1247–1256

    Article  CAS  PubMed  Google Scholar 

  • Goo JH, Park AR, Park WJ, Park OK (1999) Selection of Arabidopsis genes encoding secreted and plasma membrane proteins. Plant Mol Biol 41:415–423

    Article  CAS  PubMed  Google Scholar 

  • Grebe M, Xu J, Scheres B (2001) Cell axiality and polarity in plants—adding pieces to the puzzle. Curr Opin Plant Biol 4:520–526

    Article  CAS  PubMed  Google Scholar 

  • Hable WE, Kropf DL (1998) Roles of secretion and the cytoskeleton in cell adhesion and polarity establishment in Pelvetia compressa zygotes. Dev Biol 198:45–56

    Article  CAS  PubMed  Google Scholar 

  • Hable WE, Kropf DL (2000) Sperm entry induces polarity in fucoid zygotes. Development 127:493–501

    CAS  PubMed  Google Scholar 

  • He Z-H, He D, Kohorn BD (1998) Requirement for the induced expression of a cell wall associated receptor kinase for survival during the pathogen response. Plant J 14:55–63

    CAS  PubMed  Google Scholar 

  • He Z-H, Cheeseman I, He D, Kohorn BD (1999) A cluster of five cell wall-associated receptor kinase genes, Wak1-5, are expressed in specific organs of Arabidopsis. Plant Mol Biol 39:1189–1196

    Article  CAS  PubMed  Google Scholar 

  • Helleboid S, Chapman A, Hendriks T, Inze D, Vasseur J, Hilbert JL (2000) Cloning of beta-1,3-glucanases expressed during Cichorium somatic embryogenesis. Plant Mol Biol 42:377–386

    Article  CAS  PubMed  Google Scholar 

  • Ito H, Fukuda Y, Murata K, Kimura A (1983) Transformation of intact yeast cells treated with alkali cations. J Bacteriol 153:163–168

    CAS  PubMed  Google Scholar 

  • Jacobs KA, Collins-Racie LA, Colbert M, Duckett M, Golden-Fleet M, Kelleher K, Kriz R, LaVallie ER, Merberg D, Spaulding V, Stover J, Williamson MJ, McCoy JM (1997) A genetic selection for isolating cDNAs encoding secreted proteins. Gene 198:289–296

    Article  CAS  PubMed  Google Scholar 

  • Jinn T, Stone J, Walker J (2000) HAESA, an Arabidopsis leucine-rich repeat receptor kinase, controls floral organ abscission. Genes Dev 11:108–117

    Google Scholar 

  • Johnson AE, van Waes MA (1999) The translocon: a dynamic gateway at the ER membrane. Annu Rev Cell Dev Biol 15:799–842

    Article  PubMed  Google Scholar 

  • Kaiser CA, Preuss D, Grisafi P, Botstein D (1987) Many random sequences functionally replace the secretion signal sequence of yeast invertase. Science 235:312–317

    CAS  PubMed  Google Scholar 

  • Klein RD, Gu Q, Goddard A, Rosenthal A (1996) Selection for genes encoding secreted proteins and receptors. Proc Natl Acad Sci USA 93:7108–7113

    Article  CAS  PubMed  Google Scholar 

  • Kropf DL (1997) Induction of polarity in fucoid zygotes. Plant Cell 9:1011–1020

    Article  CAS  PubMed  Google Scholar 

  • Lally D, Ingmire P, Tong H-Y, He Z-H (2001) Antisense expression of a cell wall-associated protein kinase, WAK4, inhibits cell elongation and alters morphology. Plant Cell 13:1317–1332

    Article  CAS  PubMed  Google Scholar 

  • Li J, Chory J (1997) A putative leucine-rich repeat receptor kinase involved in brassinosteroid signal transduction. Cell 90:929–938

    PubMed  Google Scholar 

  • Li J, Wen J, Lease KA, Doke JT, Tax FE, Walker JC (2002) BAK1, an Arabidopsis LRR receptor-like protein kinase, interacts with BRI1 and modulates brassinosteroid signaling. Cell 110:213–222

    CAS  PubMed  Google Scholar 

  • Li Z, Wurtzel ET (1998) The ltk gene family encodes novel receptor-like kinases with temporal expression in developing maize endosperm. Plant Mol Biol 37:749–761

    Article  CAS  PubMed  Google Scholar 

  • Martoglio B, Dobberstein B (1998) Signal sequences: more than just greasy peptides. Trends Cell Biol 8:410–415

    CAS  PubMed  Google Scholar 

  • Nakai K, Kanehisa M (1992) A knowledge base for predicting protein localization sites in eukaryotic cells. Genomics 14:897–911

    CAS  PubMed  Google Scholar 

  • Nam KH, Li J (2002) BRI1/BAK1, a receptor kinase pair mediating brassinosteroid signaling. Cell 110:203–212

    CAS  PubMed  Google Scholar 

  • Ngsee JK, Hansen W, Walter P, Smith M (1989) Cassette mutagenic analysis of the yeast invertase signal peptide: effects on protein translocation. Mol Cell Biol 9:3400–3410

    CAS  PubMed  Google Scholar 

  • Nielsen H, Engelbrecht J, Brunak S, von Heijne G (1997) Identification of prokaryotic and eukaryotic signal peptides and prediction of their cleavage sites. Protein Eng 10:1–6

    CAS  Google Scholar 

  • Novotny AM, Forman M (1974) The relationship between changes in cell wall composition and the establishment of polarity in Fucus embryos. Dev Biol 40:162–173

    CAS  PubMed  Google Scholar 

  • Peles E, Salzer JL (2000) Molecular domains of myelinated axons. Curr Opin Neurobiol 10:558–565

    CAS  PubMed  Google Scholar 

  • Pihakaski-Maunsbach K, Griffith M, Antikainen M, Maunsbach AB (1996) Immunogold localization of glucanase-like antifreeze protein in cold acclimated winter rye. Protoplasma 191:115–125

    CAS  Google Scholar 

  • Pu R.S, Wozniak M, Robinson KR (2000) Cortical actin filaments form rapidly during photopolarization and are required for the development of calcium gradients in Pelvetia compressa zygotes. Dev Biol 222:440–449

    Article  CAS  PubMed  Google Scholar 

  • Qadota H, Python CP, Inoue SB, Arisawa M, Anraku Y, Zheng Y, Watanabe T, Levin DE, Ohya Y (1996) Identification of yeast Rho1p GTPase as a regulatory subunit of 1,3-beta-glucan synthase. Science 272:279–281

    CAS  PubMed  Google Scholar 

  • Quatrano RS (1982) Cell-wall formation in Fucus zygotes: a model system to study the assembly and localization of wall polymers. In: Brown RM (ed) Cellulose and other natural polymer systems. Plenum, London, pp 45–59

  • Quatrano RS (1997) Cortical asymmetries direct the establishment of cell polarity and the plane of cell division in the Fucus embryo. Cold Spring Harbor Symp Quant Biol 62:65–70

    CAS  PubMed  Google Scholar 

  • Quatrano RS, Crayton MA (1973) Sulfation of fucoidan in Fucus embryos I: possible role in localization. Dev Biol 30:29–35

    CAS  PubMed  Google Scholar 

  • Quatrano RS, Shaw S (1997) Role of the cell wall in the determination of cell polarity and the plane of cell division in Fucus embryos. Trends Plant Sci 2:15–21

    Article  Google Scholar 

  • Rao Z, Handford P, Mayhew M, Knott V, Brownlee GG, Stuart D (1995) The structure of a Ca2+-binding epidermal growth factor-like domain: its role in protein-protein interactions. Cell 82:131–141

    CAS  PubMed  Google Scholar 

  • Shaw SL, Quatrano RS (1996) Polar localization of a dihydropyridine receptor on living Fucus zygotes. J Cell Sci 109:335–342

    CAS  PubMed  Google Scholar 

  • Shiu SH, Bleecker AB (2001) Receptor-like kinases from Arabidopsis form a monophyletic gene family related to animal receptor kinases. Proc Natl Acad Sci USA 98:10763–10768

    Article  CAS  PubMed  Google Scholar 

  • Simmons CR (1994) The physiology and molecular biology of plant 1,3-β-d-glucanases and 1,3;1,4-β-d-glucanases. Crit Rev Plant Sci 13:325–387

    CAS  Google Scholar 

  • Singson A, Mercer KB, L'Hernault SW (1998) The C. elegans spe-9 gene encodes a sperm transmembrane protein that contains EGF-like repeats and is required for fertilization. Cell 93:71–79

    CAS  PubMed  Google Scholar 

  • Stevens PT, Quatrano RS (1978) Cell wall assembly in Fucus zygotes. II. Cellulose synthesis and deposition is controlled at the post-translational level. Dev Biol 62:518–525

    CAS  PubMed  Google Scholar 

  • Strutt D, Johnson R, Cooper K, Bray S (2002) Asymmetric localization of frizzled and the determination of notch-dependent cell fate in the Drosophila eye. Curr Biol 12:813–824

    Article  CAS  PubMed  Google Scholar 

  • Torii KU (2000) Receptor kinase activation and signal transduction in plants: an emerging picture. Curr Opin Plant Biol 3:361–367

    Article  CAS  PubMed  Google Scholar 

  • Trotochaud AE, Jeong S, Clark SE (2000) CLAVATA3, a multimeric ligand for the CLAVATA1 receptor-kinase. Science 289:613–617

    Article  CAS  PubMed  Google Scholar 

  • Vitale A, Denecke J (1999) The endoplasmic reticulum—gateway of the secretory pathway. Plant Cell 11:615–628

    CAS  PubMed  Google Scholar 

  • von Heijne G (1986) A new method for predicting signal sequence cleavage sites. Nucleic Acids Res. 14:4683–4690

  • Wagner VT, Brian L, Quatrano RS (1992) Role of vitronectin-like molecule in embryo adhesion of the brown alga Fucus. Proc Natl Acad Sci USA 89:3644–3648

    CAS  PubMed  Google Scholar 

Download references

Acknowlegements

We thank Crispin Taylor (University of North Carolina, Chapel Hill, NC, USA) for constructing the original Fucus λ cDNA library, John Fowler (Oregon State University, Corvallis, OR, USA) and Punita Nagpal (University of North Carolina, Chapel Hill, NC, USA) for helpful advice and insightful discussions, Bruce Kohorn and Zheng-Hui He (Duke University, Durham, NC, USA) for WAK2 plasmids, Richard Meagher (University of Georgia, Athens, GA, USA) for actin sequences, and the Genetics Institute (Boston, MA, USA) for the SST plasmid. This work was supported by NIH fellowship GM-18400-02 to K.D.B. and NSF grant IBN 96-04672 to R.S.Q.

Author information

Author notes

  1. Kenneth D. Belanger

    Present address: Department of Biology, Colgate University, Hamilton, NY 13346, USA

  2. Aaron J. Wyman

    Present address: Department of Molecular, Cellular, and Developmental Biology, University of Michigan, Ann Arbor, MI 48109, USA

  3. Michelle N. Sudol

    Present address: Philadelphia College of Osteopathic Medicine, Philadelphia, PA 19131, USA

  4. Sneh L. Singla-Pareek

    Present address: International Center for Genetic Engineering & Biotechnology, New Delhi, India

  5. Ralph S. Quatrano

    Present address: Department of Biology, Washington University, St. Louis, MO 63130, USA

Authors and Affiliations

  1. Department of Biology, University of North Carolina — Chapel Hill, Chapel Hill, NC 27759, USA

    Kenneth D. Belanger, Aaron J. Wyman, Sneh L. Singla-Pareek & Ralph S. Quatrano

  2. Department of Biology, Colgate University, Hamilton, NY 13346, USA

    Kenneth D. Belanger

  3. Department of Biology, University of Scranton, Scranton, PA 18510, USA

    Kenneth D. Belanger & Michelle N. Sudol

Authors
  1. Kenneth D. Belanger
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Aaron J. Wyman
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Michelle N. Sudol
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Sneh L. Singla-Pareek
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Ralph S. Quatrano
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Kenneth D. Belanger.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Belanger, K.D., Wyman, A.J., Sudol, M.N. et al. A signal peptide secretion screen in Fucus distichus embryos reveals expression of glucanase, EGF domain-containing, and LRR receptor kinase-like polypeptides during asymmetric cell growth. Planta 217, 931–950 (2003). https://doi.org/10.1007/s00425-003-1058-8

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00425-003-1058-8

Keywords

Search

Navigation