1887

Abstract

The cAMP signal transduction pathway controls many processes in fungi. The gene, encoding the regulatory subunit (PKA-R) of cAMP-dependent protein kinase (PKA), was cloned from the industrially important filamentous fungus . To investigate the involvement of PKA in morphology of , a set of transformants which overexpressed or (encoding the catalytic subunit of PKA) either individually or simultaneously was prepared as well as mutants in which and/or were disrupted. Strains overexpressing or both and could not be distinguished from the wild-type, suggesting that regulation of PKA activity is normal in these strains. Absence of PKA activity resulted in a two- to threefold reduction in colony diameter on plates. The most severe phenotype was observed in the absence of PKA-R, i.e., very small colonies on plates, absence of sporulation and complete loss of growth polarity during submerged growth. Suppressor mutations easily developed in the Δ mutant and one of these mutants appeared to lack PKA-C activity. These data suggest that cAMP-dependent protein phosphorylation in regulates growth polarity and formation of conidiospores.

Loading

Article metrics loading...

/content/journal/micro/10.1099/00221287-148-8-2635
2002-08-01
2020-04-07
Loading full text...

Full text loading...

/deliver/fulltext/micro/148/8/1482635a.html?itemId=/content/journal/micro/10.1099/00221287-148-8-2635&mimeType=html&fmt=ahah

References

  1. Adachi K., Hamer J. E. 1998; Divergent cAMP signalling pathways regulate growth and pathogenesis in the rice blast fungus Magnaporthe grisea . Plant Cell10:1361–1373[CrossRef]
    [Google Scholar]
  2. Benc̆ina M., Legis̆a M. 1999; Non-radioactive multiple reverse transcription-PCR method used for low abundance mRNA quantification. Biotechnol Tech13:865–869[CrossRef]
    [Google Scholar]
  3. Benc̆ina M., Legis̆a M. 2000; Stress mediated changes in expression of the pka C gene, encoding the catalytic subunit of cAMP-dependent protein kinase,in Aspergillus niger. Pflueg Arch439:R76–R78[CrossRef]
    [Google Scholar]
  4. Benc̆ina M., Panneman H., Ruijter G. J. G., Legis̆a M., Visser J. 1997; Characterization and overexpression of the Aspergillus niger gene encoding the cAMP-dependent protein kinase catalytic subunit. Microbiology143:1211–1220[CrossRef]
    [Google Scholar]
  5. Borges-Walmsley M. I., Walmsley A. R. 2000; cAMP signalling in pathogenic fungi: control of dimorphic switching and pathogenicity. Trends Microbiol8:133–141[CrossRef]
    [Google Scholar]
  6. Bos C. J., Debets A. J. M., Swart K., Huybers A., Kobus G., Slakhorst S. M. 1988; Genetic analysis and the construction of master strains for assignment of genes to six linkage groups in Aspergillus niger . Curr Genet14:437–443[CrossRef]
    [Google Scholar]
  7. Boy-Marcotte E., Perrot M., Bussereau F., Boucherie H., Jacquet M. 1998; Msn2p and Msn4p control large number of genes induced at the diauxic transition which are repressed by cyclic AMP in Saccharomyces cerevisiae . J Bacteriol180:1044–1052
    [Google Scholar]
  8. Bruno K. S., Aramayo R., Minke P. F., Metzenberg R. L., Plamann M. 1996; Loss of growth polarity and mislocalization of septa in a Neurospora mutant altered in the regulatory subunit of cAMP-dependent protein kinase. EMBO J15:5772–5782
    [Google Scholar]
  9. Carlson G. L., Nelson D. L. 1996; The 44-kDa regulatory subunit of the Paramecium cAMP-dependent protein kinase lacks a dimerization domain and may have a unique autophosphorylation site sequence. J Eukaryot Microbiol43:347–356[CrossRef]
    [Google Scholar]
  10. d’Enfert C. 1997; Fungal spore germination: insights from the molecular genetics of Aspergillus nidulans and Neurospora crassa . Fungal Genet Biol21:163–172[CrossRef]
    [Google Scholar]
  11. d’Enfert C., Bonini B. M., Zapella P. D., Fontaine T., da Silva A. M., Terenzi H. F. 1999; Neutral trehalases catalyse intracellular trehalose breakdown in the filamentous fungi Aspergillus nidulans and Neurospora crassa . Mol Microbiol32:471–483[CrossRef]
    [Google Scholar]
  12. Daniel P. B., Walker W. H., Habener J. F. 1998; Cyclic AMP signalling and gene regulation. Annu Rev Nutr18:353–383[CrossRef]
    [Google Scholar]
  13. de Graaff L., van den Broeck H., Visser J. 1988; Isolation and expression of the Aspergillus nidulans pyruvate kinase gene. Curr Genet13:315–321[CrossRef]
    [Google Scholar]
  14. de Oliveira J. C. F., Borges A. C. C., Marques M. V., Gomes S. L. 1997; Cloning and characterisation of the gene for the catalytic subunit of cAMP-dependent protein kinase in the aquatic fungus Blastocladiella emersonii . Eur J Biochem219:555–562
    [Google Scholar]
  15. de Voti J., Seydoux G., Beach D., McLeod M. 1991; Interaction between ran1+ protein kinase and cAMP dependent protein kinase as negative regulators of fission yeast meiosis. EMBO J10:3759–3768
    [Google Scholar]
  16. Evans M. J., Scarpulla R. C. 1988; Both upstream and intron sequence elements are required for elevated expression of the rat somatic cytochrome c gene in COS-1 cells. Mol Cell Biol8:35–41
    [Google Scholar]
  17. Fernandez M., Cao J., Villamarin J. A. 1998; In vivo phosphorylation of phosphofructokinase from the bivalve mollusc Mytilus galloprovincialis . Arch Biochem Biophys353:251–256[CrossRef]
    [Google Scholar]
  18. Frederick G. D., Rombouts P., Buxton F. P. 1993; Cloning and characterisation of pepC , a gene encoding a serine protease from Aspergillus niger . Gene125:57–64[CrossRef]
    [Google Scholar]
  19. Gancedo J. M., Mazon M. J., Eraso P. 1985; Biological role of cyclic AMP: similarities and differences between organisms. Trends Biochem Sci10:210–212[CrossRef]
    [Google Scholar]
  20. Gold S., Duncan G., Barrett K., Kronstad J. 1994; cAMP regulates morphogenesis in the fungal pathogen Ustilago maydis . Gen Dev8:2805–2816[CrossRef]
    [Google Scholar]
  21. Goncalves P. M., Griffioen G., Bebelman J. P., Planta R. J. 1997; Signalling pathway leading to transcriptional regulation of genes involved in the activation of glycolysis in yeast. Mol Microbiol25:483–493[CrossRef]
    [Google Scholar]
  22. Goosen T., Bloemheuvel G., Gysler C., de Bie D. A., van den Broek H. W., Swart K. 1987; Transformation of Aspergillus niger using the homologous orotidine-5′-phosphate-decarboxylase gene. Curr Genet11:499–503[CrossRef]
    [Google Scholar]
  23. Gradis̆nik-Grapulin M., Legis̆a M. 1997; A spontaneous change in the intracellular cyclic AMP level in Aspergillus niger is influenced by the sucrose concentration in the medium and by light. Appl Environ Microbiol63:2844–2849
    [Google Scholar]
  24. Jiang Y., Davis C., Broach J. R. 1998; Efficient transition to growth on fermentable carbon source in Saccharomyces cerevisiae requires signalling through the Ras pathway. EMBO J17:6942–6951[CrossRef]
    [Google Scholar]
  25. Johnstone I. L., Hughes J., Clutterbuck A. J. 1985; Cloning an Aspergillus nidulans developmental gene by transformation. EMBO J4:1307–1311
    [Google Scholar]
  26. Kreegipuu A., Blom N., Brunak S. 1999; PhosphoBase, a database of phosphorylation sites: release 2.0. Nucleic Acids Res7:237–239
    [Google Scholar]
  27. Kronstad J., de Maria A., Funnell D., Laidlaw R. D., Lee N., de Sa M. M., Ramesh M. 1998; Signalling via cAMP in fungi: interconnections with mitogen-activated protein kinase pathway. Arch Microbiol170:395–404[CrossRef]
    [Google Scholar]
  28. Kunisawa R., Davis T. N., Urdea M. S., Thorner J. 1987; Complete nucleotide sequence of the gene encoding the regulatory subunit of 3′, 5′-cyclic AMP-dependent protein kinase from the yeast Saccharomyces cerevisiae. Nucleic Acids Res15:368–369[CrossRef]
    [Google Scholar]
  29. Kusters-van Someren M. A., Harmsen J. A. M., Kester H. C. M., Visser J. 1991; Structure of the Aspergillus niger pelA gene and its expression in the Aspergillus niger and Aspergillus nidulans . Curr Genet20:293–299[CrossRef]
    [Google Scholar]
  30. Legis̆a M., Benc̆ina M. 1994; Evidence for the activation of 6-phosphofructo-1-kinase by cAMP-dependent protein kinase in Aspergillus niger . FEMS Microbiol Lett118:327–334[CrossRef]
    [Google Scholar]
  31. Legis̆a M., Gradis̆nik-Grapulin M. 1995; Sudden substrate dilution induces a higher rate of citric acid production by Aspergillus niger . Appl Environ Microbiol61:2732–2737
    [Google Scholar]
  32. Legis̆a M., Cimerman A., Sterle M. 1981; Germination of Aspergillus niger in a high citric acid yielding medium. FEMS Microbiol Lett11:149–152[CrossRef]
    [Google Scholar]
  33. Leon D. A., Herberg F. W., Banky P., Taylor S. S. 1997; A stable alpha-helical domain at the N terminus of the RIalpha subunits of cAMP-dependent protein kinase is a novel dimerization/docking motif. J Biol Chem272:28431–28437[CrossRef]
    [Google Scholar]
  34. Maeda T., Watanabe Y., Kunitomo H., Yamamoto M. 1994; Cloning of pka1 gene encoding the catalytic subunit of the cAMP-dependent protein kinase in Schizosaccharomyces pombe. J Biol Chem269:9632–9637
    [Google Scholar]
  35. Marques M. V., Gomes L. S. 1992; Cloning and structural analysis of the gene for the regulatory subunit of cAMP-dependent protein kinase in Blastocladiella emersonii . J Biol Chem267:17201–17207
    [Google Scholar]
  36. Mitchell T. K., Dean R. A. 1995; The cAMP-dependent protein kinase catalytic subunit is required for appressorium formation and pathogenesis by the rice blast pathogen Magnaporthe grisea . Plant Cell7:1869–1878[CrossRef]
    [Google Scholar]
  37. Murre R., Schonheber P., Baltimore D. 1989; A new DNA-binding and dimerization motif in immunoglobulin enhancer binding, daughterless, MyoD and myc proteins. Cell56:777–783[CrossRef]
    [Google Scholar]
  38. Mutzel R., Lacombe M.-L., Simon M.-N., de Gunzburg J., Veron M. 1987; Cloning and cDNA sequence of the regulatory subunit of cAMP-dependent protein kinase from Dictyostelium discoideum . Proc Natl Acad Sci USA84:6–10[CrossRef]
    [Google Scholar]
  39. Niimi M. 1996; Dibutyryl cyclic AMP-enhanced germ tube formation in exponentially growing Candida albicans cells. Fungal Genet Biol20:79–83[CrossRef]
    [Google Scholar]
  40. Orlowski M. 1991; Mucor dimorphism. Microbiol Rev55:234–258
    [Google Scholar]
  41. Pontecorvo G., Roper J. A., Hemmons L. J., MacDonald K. D., Bufton A. W. J. 1953; The genetics of Aspergillus nidulans . Adv Genet5:141–238
    [Google Scholar]
  42. Quandt K., Frech K., Karas H., Wingender E., Werner T. 1995; MarInd and MatInspector-New fast and versatile tools for determination of consensus matches in nucleotide sequence data. Nucleic Acids Res23:4878–4884[CrossRef]
    [Google Scholar]
  43. Sambrook J., Fritsch E. F., Maniatis T. 1989; Molecular Cloning: a Laboratory Manual, 2nd edn. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory;
    [Google Scholar]
  44. Sanger F., Nicklen S., Coulson A. R. 1977; DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci USA74:5463–5467[CrossRef]
    [Google Scholar]
  45. Su Y., Dostmann W. R., Herberg F. W., Durick K., Xuong N. H., Ten Eyck L., Taylor S. S., Varughese K. I. 1995; Regulatory subunit of protein kinase A: structure of deletion mutant with cAMP binding domains. Science269:807–813[CrossRef]
    [Google Scholar]
  46. Taylor S. S., Knighton D. R., Zheng J., Eyck L. F. T., Sowadski J. M. 1992; Structural framework for the protein kinase family. Annu Rev Cell Biol8:429–462[CrossRef]
    [Google Scholar]
  47. Treger J. M., Magee T. R., McEntee K. 1998; Functional analysis of the stress response element and its role in the multistress response of Saccharomyces cerevisiae . Biochem Biophys Res Commun243:13–19[CrossRef]
    [Google Scholar]
  48. Unkles S. E. 1992; Gene organisation in industrial filamentous fungi. In Applied Molecular Genetics of Filamentous Fungi pp28–53 Edited by Kinghorn J. R., Turner G.. London: Blackie;
    [Google Scholar]
  49. Van Diepeningen A. D., Debets A. J. M., Hoekstra R. F. 1998; Intra- and interspecies virus transfer in aspergilli via protoplast fusion. Fungal Genet Biol25:171–180[CrossRef]
    [Google Scholar]
  50. Visniac W., Santer M. 1957; The thiobacili. Bacteriol Rev21:195–213
    [Google Scholar]
  51. Zahringer H., Holzer H., Nwaka S. 1998; Stability of neutral trehalase during heat stress in Saccharomyces cerevisiae is dependent on the activity of the catalytic subunit of cAMP-dependent protein kinase, Tpk1 and Tpk2. Eur J Biochem255:544–551[CrossRef]
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/micro/10.1099/00221287-148-8-2635
Loading
/content/journal/micro/10.1099/00221287-148-8-2635
Loading

Data & Media loading...

Most cited this month

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