1887

Microbiology Society logo

Volume 148, Issue 9

Suppression of fruit-body formation by constitutively active G-protein α-subunits and in the homobasidiomycete

The GenBank accession numbers for the sequences reported in this paper are AB066503, AB051903 and AB051904.

Free

Abstract

The heterotrimeric G-protein α-subunit family plays multiple roles in eukaryotic cells, such as the regulation of growth and development, of pathogenicity and of the transmission of pheromone stimulation. In the homobasidiomycete , some genes encoding heterotrimeric G-protein α-subunits (α, α, , and ) have been reported. In this study, constitutively active mutants of , and were generated by site-directed mutagenesis and introduced into the monokaryon strain to investigate the function of each gene. Northern analysis showed that the mutated genes were strongly expressed when compared with endogenous G-proteins in many clones. Upon macroscopic examination, some transformed clones expressing (Q207R) and (Q204R) mutant genes exhibited a slight suppression of aerial-hyphae formation in the monokaryon strain. In contrast to the slight suppression of aerial-hyphae formation in the monokaryon, most clones expressing mutated or genes failed to form fruit-bodies in the dikaryon strain. This observation indicated that and played a role in suppressing fruit-body formation in the dikaryon. Furthermore, these phenotypes were similar to the phenotype of the mutant in to some extent. Since the gene encodes a putative regulator of the G-protein signalling protein (RGS), and might be targets of .

  • Received:
  • Accepted:
  • Revised:
  • Published Online:
Keyword(s): GPD, glyceraldehyde-3-phosphate dehydrogenase , RGS, G-protein signalling protein , ScGP-A, Schizophyllum commune G-protein α-subunit A , ScGP-B, S. commune G-protein α-subunit B , ScGP-C, S. commune G-protein α-subunit C and site-directed mutagenesis
© Microbiology Society
Loading

Article metrics loading...

/content/journal/micro/10.1099/00221287-148-9-2797
2002-09-01
2024-04-19
Loading full text...

Full text loading...

/deliver/fulltext/micro/148/9/1482797a.html?itemId=/content/journal/micro/10.1099/00221287-148-9-2797&mimeType=html&fmt=ahah

References

  1. Barnes W. M. 1994; PCR amplification of up to 35-kb DNA with high fidelity and high yield from λ bacteriophage templates. Proc Natl Acad Sci USA 91:2216–2220 [CrossRef]
     [Google Scholar]
  2. Berman D. M., Wilkie T. M., Gilman A. G. 1996a; GAIP and RGS4 are GTPase-activating proteins for the Gi subfamily of G protein α subunits. Cell 86:445–452 [CrossRef]
     [Google Scholar]
  3. Berman D. M., Kozasa T., Gilman A. G. 1996b; The GTPase-activating protein RGS4 stabilizes the transition state for nucleotide hydrolysis. J Biol Chem 271:27209–27212 [CrossRef]
     [Google Scholar]
  4. Clackson T., Gussow D., Jones P. T. 1994; General applications of PCR to gene cloning and manipulation. In The Polymerase Chain Reaction Edited by Mullis K. B., Ferre F., Gibbs R. A. BirkHauser:
     [Google Scholar]
  5. Clapham D. E., Neer E. J. 1993; New roles for G-protein βγ-dimers in transmembrane signalling. Nature 365:403–406 [CrossRef]
     [Google Scholar]
  6. Coleman D. E., Berghuis A. M., Lee E., Linder M. E., Gilman A. G., Sprang S. R. 1994; Structures of active conformations of Giα1 and the mechanism of GTP hydrolysis. Science 265:1405–1412 [CrossRef]
     [Google Scholar]
  7. Fowler T. J., Mitton M. F. 2000; Scooter, a new active transposon in Schizophyllum commune , has disrupted two genes regulating signal transduction. Genetics 156:1585–1594
     [Google Scholar]
  8. Gola S., Hegner J., Kothe E. 2000; Chimeric pheromone receptors in the basidiomycete Schizophyllum commune . Fungal Genet Biol 30:191–196 [CrossRef]
     [Google Scholar]
  9. Hamm H. E. 1998; The many faces of G protein signaling. J Biol Chem 273:669–672 [CrossRef]
     [Google Scholar]
  10. Hanahan D. 1983; Studies on transformation of Escherichia coli with plasmids. J Mol Biol 166:557–580 [CrossRef]
     [Google Scholar]
  11. Harmsen M. C., Schuren F. H., Moukha S. M., van Zuilen C. M., Punt P. J., Wessels J. G. 1992; Sequence analysis of the glyceraldehyde-3-phosphate dehydrogenase genes from the basidiomycetes Schizophyllum commune , Phanerochaete chrysosporium and Agaricus bisporus . Curr Genet 22:447–454 [CrossRef]
     [Google Scholar]
  12. Hepler J. R., Berman D. M., Gilman A. G., Kozasa T. 1997; RGS4 and GAIP are GTPase-activating proteins for Gqα and block activation of phospholipase Cβ by γ-thio-GTP-Gqα . Proc Natl Acad Sci USA 94:428–432 [CrossRef]
     [Google Scholar]
  13. Isshiki T., Mochizuki N., Maeda T., Yamamoto M. 1992; Characterization of a fission yeast gene, gpa2 , that encodes a Gα subunit involved in the monitoring of nutrition. Genes Dev 6:2455–2462 [CrossRef]
     [Google Scholar]
  14. Kothe E. 1996; Tetrapolar fungal mating types: sexes by the thousands. FEMS Microbiol Rev 18:65–87 [CrossRef]
     [Google Scholar]
  15. Kothe E. 1999; Mating types and pheromone recognition in the Homobasidiomycete Schizophyllum commune . Fungal Genet Biol 27:146–152 [CrossRef]
     [Google Scholar]
  16. Kozak K. R., Ross I. K. 1991; Signal transduction in Coprinus congregatus : evidence for the involvement of G proteins in blue light photomorphogenesis. Biochem Biophys Res Comm 179:1225–1231 [CrossRef]
     [Google Scholar]
  17. Kozak K. R., Foster L. M., Ross I. K. 1995; Cloning and characterization of a G protein α-subunit-encoding gene from the basidiomycete, Coprinus congregatus . Gene 163:133–137 [CrossRef]
     [Google Scholar]
  18. Kubler E., Mosch H. U., Rupp S., Lisanti M. P. 1997; Gpa2p, a G-protein α-subunit, regulates growth and pseudohyphal development in Saccharomyces cerevisiae via a cAMP-dependent mechanism. J Biol Chem 272:20321–20323 [CrossRef]
     [Google Scholar]
  19. Kurjan J. 1993; The pheromone response pathway in Saccharomyces cerevisiae . Annu Rev Genet 27:147–179 [CrossRef]
     [Google Scholar]
  20. Landis C. A., Masters S. B., Spada A., Pace A. M., Bourne H. R., Vallar L. 1989; GTPase inhibiting mutations activate the α chain of Gs and stimulate adenylyl cyclase in human pituitary tumours. Nature 340:692–696 [CrossRef]
     [Google Scholar]
  21. Leberer E., Dignard D., Thomas D. Y., Leeuw T. 2000; A conserved Gβ binding (GBB) sequence motif in Ste20p/PAK family protein kinases. Biol Chem 381:427–431
     [Google Scholar]
  22. Liu S., Dean R. A. 1997; G protein α subunit genes control growth, development, and pathogenicity of Magnaporthe grisea . Mol Plant–Microbe Interact 10:1075–1086 [CrossRef]
     [Google Scholar]
  23. Liu Y. G, Whittier R. F. 1995; Thermal asymmetric interlaced PCR: automatable amplification and sequencing of insert end fragments from P1 and YAC clones for chromosome walking. Genomics 25:674–681 [CrossRef]
     [Google Scholar]
  24. Marchuk D., Drumm M., Saulino A., Collins F. S. 1991; Construction of T-vectors, a rapid and general system for direct cloning of unmodified PCR products. Nucleic Acids Res 19:1154 [CrossRef]
     [Google Scholar]
  25. McPherson M. J., Jones K. M., Gurr S. J. 1994; PCR with highly degenerate primers. In The Polymerase Chain Reaction Edited by Mullis K. B., Ferre F., Gibbs R. A. BirkHauser:
     [Google Scholar]
  26. Miyajima I., Nakafuku M., Nakayama N., Brenner C., Miyajima A., Kaibuchi K., Arai K., Kaziro Y., Matsumoto K. 1987; GPA1 , a haploid-specific essential gene, encodes a yeast homolog of mammalian G protein which may be involved in mating factor signal transduction. Cell 50:1011–1019 [CrossRef]
     [Google Scholar]
  27. Morris A. J., Malbon C. C. 1999; Physiological regulation of G-protein-linked signaling. Physiol Rev 80:1373–1430
     [Google Scholar]
  28. Munoz-Rivas A. M., Specht C. A., Ullrich R. C., Novotny C. P. 1986a; Isolation of the DNA sequence coding indole-3-glycerol phosphate synthetase and phosphoribosylanthranilate isomerase of Schizophyllum commune . Curr Genet 10:909–913 [CrossRef]
     [Google Scholar]
  29. Munoz-Rivas A., Specht C. A., Drummond B. J., Froeliger E., Novotny C. P., Ullrich R. C. 1986b; Transformation of the basidiomycete, Schizophyllum commune . Mol Gen Genet 205:103–106 [CrossRef]
     [Google Scholar]
  30. Murakami M., Kamiya Y., Yanagita Y., Mori M. 1999; Gsα mutations in hyperfunctioning thyroid adenomas. Arch Med Res 30:514–521 [CrossRef]
     [Google Scholar]
  31. Perkins J. H. 1969; Morphogenesis in Schizophyllum commune . I. Effects of white light. Plant Physiol 44:1706–1711 [CrossRef]
     [Google Scholar]
  32. Perkins J. H., Gordon S. A. 1969; Morphogenesis in Schizophyllum commune . II. Effects of monochromatic light. Plant Physiol 44:1712–1716 [CrossRef]
     [Google Scholar]
  33. Raper C. A. 1988; Schizophyllum commune , a model for genetic studies of the Basidiomycotina. In Advances in Plant Pathology pp 511–522 Edited by King R. C. New York: Plenum Press;
     [Google Scholar]
  34. Raper J. R., Miles P. G. 1958; The genetics of Schizophyllum commune . Genetics 43:530–546
     [Google Scholar]
  35. Regenfelder E., Spellig T., Hartmann A., Lauenstein S., Bolker M., Kahmann R. 1997; G proteins in Ustilago maydis : transmission of multiple signals?. EMBO J 16:1934–1942 [CrossRef]
     [Google Scholar]
  36. Schuren F. H., Wessels J. G. 1990; Two genes specifically expressed in fruiting dikaryons of Schizophyllum commune : homologies with a gene not regulated by mating-type genes. Gene 90:199–205 [CrossRef]
     [Google Scholar]
  37. Schuurs T. A., Schaeffer E. A., Wessels J. G. 1997; Homology-dependent silencing of the SC3 gene in Schizophyllum commune . Genetics 147:589–596
     [Google Scholar]
  38. Schwalb M. N., Miles P. G. 1967; Morphogenesis of Schizophyllum commune . II. Effect of microaerobic growth. Mycologia 59:610–622 [CrossRef]
     [Google Scholar]
  39. Specht C. A. 1995; Isolation of the Bα and Bβ mating-type loci of Schizophyllum commune . Curr Genet 28:374–379 [CrossRef]
     [Google Scholar]
  40. Uno I., Ishikawa T. 1973; Purification and identification of the fruiting-inducing substances in Coprinus macrorhizus . J Bacteriol 113:1240–1248
     [Google Scholar]
  41. Vaillancourt L. J., Raudaskoski M., Specht C. A., Raper C. A. 1997; Multiple genes encoding pheromones and a pheromone receptor define the Bβ1 mating-type specificity in Schizophyllum commune . Genetics 146:541–551
     [Google Scholar]
  42. van Wetter M. A., Wosten H. A., Sietsma J. H., Wessels J. G. 2000; Hydrophobin gene expression affects hyphal wall composition in Schizophyllum commune . Fungal Genet Biol 31:99–104 [CrossRef]
     [Google Scholar]
  43. Wendland J., Vaillancourt L. J., Hegner J., Lengeler K. B., Laddison K. J., Specht C. A., Raper C. A., Kothe E. 1995; The mating-type locus Bα1 of Schizophyllum commune contains a pheromone receptor gene and putative pheromone genes. EMBO J 14:5271–5278
     [Google Scholar]
  44. Wessels J. G. 1997; Hydrophobins: proteins that change the nature of the fungal surface. Adv Microb Physiol 38:1–45
     [Google Scholar]
  45. Wessels J. G., de Vries O. M., Asgeirsdottir S. A., Springer J. 1991; The thn mutation of Schizophyllum commune , which suppresses formation of aerial hyphae, affects expression of the SC3 hydrophobin gene. J Gen Microbiol 137:2439–2445 [CrossRef]
     [Google Scholar]
  46. Wösten H. A., Asgeirsdottir S. A., Krook J. H., Drenth J. H., Wessels J. G. 1994; The fungal hydrophobin SC3p self-assembles at the surface of aerial hyphae as a protein membrane constituting the hydrophobic rodlet layer. Eur J Cell Biol 63:122–129
     [Google Scholar]
  47. Yu J. H., Wieser J., Adams T. H. 1996; The Aspergillus FlbA RGS domain protein antagonizes G protein signaling to block proliferation and allow development. EMBO J 15:5184–5190
     [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/micro/10.1099/00221287-148-9-2797
Loading
Suppression of fruit-body formation by constitutively active G-protein α-subunits ScGP-A and ScGP-C in the homobasidiomycete Schizophyllum communeThe GenBank accession numbers for the sequences reported in this paper are AB066503, AB051903 and AB051904.
Microbiology 148, 2797 (2002); https://doi.org/10.1099/00221287-148-9-2797
/content/journal/micro/10.1099/00221287-148-9-2797
/content/journal/micro/10.1099/00221287-148-9-2797
Loading

Data & Media loading...

Most cited Most Cited RSS feed

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error