Advertisement

Current Genetics

, Volume 51, Issue 6, pp 367–375 | Cite as

The fruiting-specific Le.flp1 gene, encoding a novel fungal fasciclin-like protein, of the basidiomycetous mushroom Lentinula edodes

  • Yasumasa MiyazakiEmail author
  • Shinya Kaneko
  • Masahide Sunagawa
  • Kazuo Shishido
  • Takashi Yamazaki
  • Masaya Nakamura
  • Katsuhiko Babasaki
Research Article

Abstract

To understand the molecular mechanisms of fruiting body formation of basidiomycetous mushrooms, we have isolated over a 100 of developmentally regulated genes that were specifically transcribed during fruiting body development in Lentinula edodes (Shiitake-mushroom) by a subtractive hybridization, cDNA-RDA (cDNA representational difference analysis). One of these genes, named Le.flp1, was isolated from the primordial cDNA library of L. edodes, and the expression product of Le.flp1 and putative fungal homologues contained a characteristic region, homologous to the Fas domain of fasciclin family proteins, which are capable of promoting cell adhesion through Fas domain-mediated homophilic interactions in various organisms. RT-PCR analyses suggested that Le.flp1 was specifically expressed in primordia and mature fruiting bodies. In situ hybridization indicated that Le.flp1 transcripts were distributed distinctly in the following tissues: the inside of gills of fruiting bodies, especially at the boundary between the subhymenium and trama, where there is active proliferation of basidium cells for producing basidiospores; peripheral regions of the primordium, pileus and stipe; and both inner tissue and outer regions of the stipe. Our results suggest the hypothesis that Le.flp1 plays a role in cellular differentiation and development in ubiquitous tissues during fruiting body formation in L. edodes, possibly through cell adhesion.

Keywords

Basidiomycete Mushroom Lentinula edodes Fruiting body formation In situ hybridization Fasciclin 

Notes

Acknowledgments

This work was supported by a Grant-in-Aid for Scientific Research (No. 17780068) from the Ministry of Education, Culture, Sports, Science and Technology of Japan. The authors deeply appreciate Dr. Teruko Konishi, of the University of Ryukyus’ Faculty of Agriculture, for her useful and accurate comments on the experiments.

References

  1. Altschul SF, Madden TL, SchaVer AA, Zhang J, Zhang Z, Miller W, Lipman DJ (1997) Gapped BLAST and PSI-BLAST: a new generation of protein database search programs. Nucleic Acids Res 25:3389–3402PubMedCrossRefGoogle Scholar
  2. Bairoch A, Bucher P, Hofmann K (1997) The PROSITE database, its status in 1997. Nucleic Acids Res 25:217–221PubMedCrossRefGoogle Scholar
  3. Carr MD, Bloemink MJ, Dentten E, Whelan AO, Gordon SV, Kelly G, Frenkiel TA, Hewinson RG, Williamson RA (2003) Solution structure of the Mycobacterium tuberculosis complex protein MPB70: from tuberculosis pathogenesis to inherited human corneal desease. J Biol Chem 278:43736–43743PubMedCrossRefGoogle Scholar
  4. Chang ST, Miles PG (1991) Recent trends in world production of cultivated mushrooms. Mush J 503:15–18Google Scholar
  5. Chiu SW, Moore D (1990) Sporulation in Coprinus cinereus: use of an in vitro assay to establish the major landmarks in differentiation. Mycol Res 94:249–253CrossRefGoogle Scholar
  6. Dahiya P, Findlay K, Roberts K, McCann MC (2006) A fasciclin-domain containing gene, ZeFLA11, is expressed exclusively in xylem elements that have reticulate wall thickenings in the stem vascular system of Zinnia elegans cv Envy. Planta 223:1281–1291PubMedCrossRefGoogle Scholar
  7. Eisenhaber B, Wildpaner M, Schultz CJ, Borner GHH, Dupree P, Eisenhaber F (2003) Glycosylphosphatidylinositol lipid anchoring of plant proteins: sensitive prediction from sequence- and genome-wide studies for Arabidopsis and rice. Plant Physiol 133:1691–1701PubMedCrossRefGoogle Scholar
  8. Elkins T, Hortsch M, Bieber AJ, Snow PM, Goodman CS (1990a) Drosophila fasciclin I is a novel homophilic adhesion molecule that along with fasciclin III can mediate cell sorting. J Cell Biol 110:1825–1832PubMedCrossRefGoogle Scholar
  9. Elkins T, Zinn K, McAllister L, Hoffmann FM, Goodman CS (1990b) Genetic analysis of a Drosophila neural cell adhesion molecule: interaction of fasciclin I and abelson tyrosine kinase mutations. Cell 60:565–575PubMedCrossRefGoogle Scholar
  10. Endo H, Kajiwara S, Tsunoka O, Shishido K (1994) A novel cDNA, priBc encoding a protein with a Zn(II)2Cys6 zinc cluster DNA-binding motif, derived from the basidiomycete Lentinus edodes. Gene 139:117–121PubMedCrossRefGoogle Scholar
  11. Faik A, Abouzouhair J, Sarhan F (2006) Putative fasciclin-like arabinogalactan-proteins (FLA) in wheat (Triticum aestivum) and rice (Oryza sativa): identification and bioinformatic analyses. Mol Genet Genomics 276:478–494PubMedCrossRefGoogle Scholar
  12. Felsenstein J (1985) Confidence limits on phylogenies: an approach using the bootstrap. Evolution 39:783–791CrossRefGoogle Scholar
  13. Fitch WM (1977) On the problem of discovering the most parsimonious tree. Am Nat 111:223–257CrossRefGoogle Scholar
  14. Galagan JE, Calvo SE, Cuomo C, Ma LJ, Wortman JR, Batzoglou S, Lee SI, Basturkmen M, Spevak CC, Clutterbuck J, Kapitonov V, Jurka J, Scazzocchio C, Farman M, Butler J, Purcell S, Harris S, Braus GH, Draht O, Busch S, D’Enfert C, Bouchier C, Goldman GH, Bell-Pedersen D, Griffiths-Jones S, Doonan JH, Yu J, Vienken K, Pain A, Freitag M, Selker EU, Archer DB, Penalva MA, Oakley BR, Momany M, Tanaka T, Kumagai T, Asai K, Machida M, Nierman WC, Denning DW, Caddick M, Hynes M, Paoletti M, Fischer R, Miller B, Dyer P, Sachs MS, Osmani SA, Birren BW (2005) Sequencing of Aspergillus nidulans and comparative analysis with A. fumigatus and A. oryzae. Nature 438:1105–1115PubMedCrossRefGoogle Scholar
  15. Gaspar YM, Johnson KL, Mckenna JA, Bacic A, Schultz CJ (2001) The complex structures of arabinogalactan-proteins and the journey towards a function. Plant Mol Biol 47:161–176PubMedCrossRefGoogle Scholar
  16. Han JH, Stratowa C, Rutter WJ (1987) Isolation of full-length putative rat lysophospholipase cDNA using improved methods for mRNA isolation and cDNA cloning. Biochemistry 26:1617–1625PubMedCrossRefGoogle Scholar
  17. Hardtke CS (2006) Root development–branching into novel spheres. Curr Opin Plant Biol 9:66–71PubMedCrossRefGoogle Scholar
  18. Hori K, Kajiwara S, Saito T, Miyazawa H, Katayose Y, Shishido K (1991) Cloning, sequence analysis and transcriptional expression of a ras gene of the edible basidiomycete Lentinus edodes. Gene 105:91–96PubMedCrossRefGoogle Scholar
  19. Hubank M, Schatz DG (1994) Identifying differences in mRNA expression by representational difference analysis of cDNA. Nucleic Acids Res 22:5640–5648PubMedCrossRefGoogle Scholar
  20. Huber O, Sumper M (1994) Algal-CAMs: isoforms of a cell adhesion molecule in embryos of the alga Volvox with homology to Drosophila fasciclin I. EMBO J 13:4212–4222PubMedGoogle Scholar
  21. Johnson KL, Jones BJ, Bacic A, Schultz CJ (2003) The fasciclin-like arabinogalactan proteins of Arabidopsis. A multigene family of putative cell adhesion molecules. Plant Physiol 133:1911–1925PubMedCrossRefGoogle Scholar
  22. Kajiwara S, Yamaoka K, Hori K, Miyazawa H, Saito T, Kanno T, Shishido K (1992) Isolation and sequence of a developmentally regulated putative novel gene, priA, from the basidiomycete Lentinus edodes. Gene 114:173–178PubMedCrossRefGoogle Scholar
  23. Kaneko S, Shishido K (2001) Cloning and sequence analysis of the basidiomycete Lentinus edodes ribonucleotide reductase small subunit cDNA and expression of a corresponding gene in L. edodes. Gene 262:43–50PubMedCrossRefGoogle Scholar
  24. Kawamoto T, Noshiro M, Shen M, Nakamasu K, Hashimoto K, Kawashima-Ohya Y, Gotoh O, Kato Y (1998) Structural and phylogenetic analyses of RGP-CAP/big-h3, a fasciclin-like adhesion protein expressed in chick chondrocytes. Biochim Biophys Acta 1395:288–292PubMedGoogle Scholar
  25. Kim JE, Kim SJ, Lee BH, Park RW, Kim KS, Kim IS (2000) Identification of motifs for cell adhesion within the repeated domains of transforming growth factor-b-induced gene, big-h3. J Biol Chem 275:30907–30915PubMedCrossRefGoogle Scholar
  26. Kondoh O, Muto A, Kajiwara S, Takagi J, Saito T, Shishido K (1995) A fruit body-specific cDNA mfbAc from the mushroom Lentinus edodes encodes a high-molecular-weight cell-adhesion protein containing an Arg-Gly-Asp motif. Gene 154:31–37PubMedCrossRefGoogle Scholar
  27. Kües U (2000) Life history and developmental processes in the basidiomycete Coprinus cinereus. Microbiol Mol Biol Rev 64:316–353PubMedCrossRefGoogle Scholar
  28. Lazdunski CJ, Benedetti H (1990) Insertion and translocation of proteins into and through membranes. FEBS Lett 268:408–414PubMedCrossRefGoogle Scholar
  29. Machida M, Asai K, Sano M, Tanaka T, Kumagai T, Terai G, Kusumoto K, Arima T, Akita O, Kashiwagi Y, Abe K, Gomi K, Horiuchi H, Kitamoto K, Kobayashi T, Takeuchi M, Denning DW, Galagan JE, Nierman WC, Yu J, Archer DB, Bennett JW, Bhatnagar D, Cleveland TE, Fedorova ND, Gotoh O, Horikawa H, Hosoyama A, Ichinomiya M, Igarashi R, Iwashita K, Juvvadi PR, Kato M, Kato Y, Kin T, Kokubun A, Maeda H, Maeyama N, Maruyama J, Nagasaki H, Nakajima T, Oda K, Okada K, Paulsen I, Sakamoto K, Sawano T, Takahashi M, Takase K, Terabayashi Y, Wortman JR, Yamada O, Yamagata Y, Anazawa H, Hata Y, Koide Y, Komori T, Koyama Y, Minetoki T, Suharnan S, Tanaka A, Isono K, Kuhara S, Ogasawara N, Kikuchi H (2005) Genome sequencing and analysis of Aspergillus oryzae. Nature 438:1157–1161PubMedCrossRefGoogle Scholar
  30. Miyazaki Y, Jojima T, Ono T, Yamazaki T, Shishido K (2004a) A cDNA homologue of Schizosaccharomyces pombe cdc5 + from the mushroom Lentinula edodes: characterization of the cDNA and its expressed product. Biochim Biophys Acta 1680:93–102PubMedGoogle Scholar
  31. Miyazaki Y, Nakamura M, Babasaki K (2005) Molecular cloning of developmentally specific genes by representational difference analysis during the fruiting body formation in the basidiomycete Lentinula edodes. Fungal Genet Biol 42:493–505PubMedCrossRefGoogle Scholar
  32. Miyazaki Y, Sakuragi Y, Yamazaki T, Shishido K (2004b) Target genes of the developmental regulator PRIB of the mushroom Lentinula edodes. Biosci Biotechnol Biochem 68:1898–1905PubMedCrossRefGoogle Scholar
  33. Miyazaki Y, Tsunoka O, Shishido K (1997) Determination of the DNA-binding sequences of the Zn(II)2Cys6 zinc-cluster-containing PRIB protein, derived from the basidiomycete Lentinus edodes gene. J Biochem 122:1088–1091PubMedGoogle Scholar
  34. Nakai K, Horton P (1999) PSORT: a program for detecting sorting signals in proteins and predicting their subcellular localization. Trends Biochem Sci 24:34–36PubMedCrossRefGoogle Scholar
  35. Nakazawa T, Miyazaki Y, Kaneko S, Shishido K (2006) Developmental regulator Le.CDC5 of the mushroom Lentinula edodes: analyses of its amount in each of the stages of fruiting-body formation and its distribution in parts of the fruiting bodies. FEMS Microbiol Lett 261:60–63PubMedCrossRefGoogle Scholar
  36. Ng WL, Ng TP, Kwan HS (2000) Cloning and characterization of two hydrophobin genes differentially expressed during fruiting body development in Lentinula edodes. FEMS Micobiol Lett 185:139–145CrossRefGoogle Scholar
  37. Nierman WC, Pain A, Anderson MJ, Wortman JR, Kim HS, Arroyo J, Berriman M, Abe K, Archer DB, Bermejo C, Bennett J, Bowyer P, Chen D, Collins M, Coulsen R, Davies R, Dyer PS, Farman M, Fedorova N, Fedorova N, Feldblyum TV, Fischer R, Fosker N, Fraser A, Garcia JL, Garcia MJ, Goble A, Goldman GH, Gomi K, Griffith-Jones S, Gwilliam R, Haas B, Haas H, Harris D, Horiuchi H, Huang J, Humphray S, Jimenez J, Keller N, Khouri H, Kitamoto K, Kobayashi T, Konzack S, Kulkarni R, Kumagai T, Lafon A, Latge JP, Li W, Lord A, Lu C, Majoros WH, May GS, Miller BL, Mohamoud Y, Molina M, Monod M, Mouyna I, Mulligan S, Murphy L, O’Neil S, Paulsen I, Penalva MA, Pertea M, Price C, Pritchard BL, Quail MA, Rabbinowitsch E, Rawlins N, Rajandream MA, Reichard U, Renauld H, Robson GD, Rodriguez de Cordoba S, Rodriguez-Pena JM, Ronning CM, Rutter S, Salzberg SL, Sanchez M, Sanchez-Ferrero JC, Saunders D, Seeger K, Squares R, Squares S, Takeuchi M, Tekaia F, Turner G, Vazquez de Aldana CR, Weidman J, White O, Woodward J, Yu JH, Fraser C, Galagan JE, Asai K, Machida M, Hall N, Barrell B, Denning DW (2005) Genomic sequence of the pathogenic and allergenic filamentous fungus Aspergillus fumigatus. Nature 438:1151–1156PubMedCrossRefGoogle Scholar
  38. Nishizawa H, Miyazaki Y, Kaneko S, Shishido K (2002) Distribution of hydrophobin 1 gene transcript in developing fruiting bodies of Lentinula edodes. Biosci Biotechnol Biochem 66:1951–1954PubMedCrossRefGoogle Scholar
  39. Paulsrud P, Lindblad P (2002) Fasciclin domain proteins are present in nostoc symbionts of lichens. Appl Environ Microbiol 68:2036–2039PubMedCrossRefGoogle Scholar
  40. Reijnders AFM (1979) Developmental anatomy of Coprinus. Persoonia 10:383–424Google Scholar
  41. Saitou N, Nei M (1987) The neighbor-joining method: a new method for reconstructing phylogenetic trees. Mol Biol Evol 4:406–425PubMedGoogle Scholar
  42. Sambrook J, Russell DW (2001) Molecular Cloning: A Laboratory Manual, 3rd edn. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NYGoogle Scholar
  43. Takagi Y, Katayose Y, Shishido K (1988) Intracellular levels of cyclic AMP and adenylated cyclase activity during mycelial development in fruit body formation in L. edodes. FEMS Microbiol Lett 55:275–278CrossRefGoogle Scholar
  44. Tanaka Y, Kaneko S, Katsukawa S, Yamazaki T, Ohta A, Miyazaki Y, Shishido K (2005) Specific distribution in homobasidiomycete hymenophores of the transcripts of Ras protein and G-protein alpha-subunit genes. FEMS Microbiol Lett 242:169–175PubMedCrossRefGoogle Scholar
  45. Tsivileva OM, Nikitina VE, Garibova LV, Ignatov VV (2001) Lectin activity of Lentinus edodes. Int Microbiol 4:41–45PubMedGoogle Scholar
  46. von Heijne G (1986) A new method for predicting signal cleavage sites. Nucleic Acids Res 14:4683–4690CrossRefGoogle Scholar

Copyright information

© Springer-Verlag 2007

Authors and Affiliations

  • Yasumasa Miyazaki
    • 1
    Email author
  • Shinya Kaneko
    • 2
  • Masahide Sunagawa
    • 1
  • Kazuo Shishido
    • 2
  • Takashi Yamazaki
    • 2
    • 3
  • Masaya Nakamura
    • 1
  • Katsuhiko Babasaki
    • 1
  1. 1.Department of Applied MicrobiologyForestry and Forest Products Research InstituteTsukuba-NorinJapan
  2. 2.Department of Life Science, Graduate School of Bioscience and BiotechnologyTokyo Institute of TechnologyMidori-kuJapan
  3. 3.Institute of Space and Astronautical ScienceJapan Aerospace Exploration AgencyTsukubaJapan

Personalised recommendations