1887

Abstract

The gene encoding the catalytic subunit of cAMP-dependent protein kinase has been isolated from the industrially important filamentous fungus A probe for screening phage libraries was generated by a polymerase chain reaction using degenerate primers. cDNA and genomic DNA clones were isolated and sequenced. An open reading frame of 1440 bp, interrupted by three short introns, encodes a polypeptide of 480 amino acids with a calculated molecular mass of 53813 Da. The cAMP-dependent protein kinase catalytic subunit (PKA-C) from has a 126 amino acid extension at the N-terminus compared to the PKA-C of higher eukaryotes that - except for the first 15 amino acids, which are homologous to the PKA-C - shows no significant similarity to the N-terminal extension of PKA-C of other lower eukaryotes. The catalytic core of PKA-C of A. shows extensive homology with the PKA-C isolated from all other eukaryotes. Low-stringency hybridization did not reveal any other homologue in The cloned was used for transformation of leading to increased levels of mRNA and PKA-C activity. Transformants overexpressing were phenotypically different with respect to growth, showing a more compact colony morphology, accompanied by a more dense sporulation, especially on media containing trehalose and glycerol. A number of transformants also showed a strongly reduced or complete absence of sporulation. This phenotype was quickly lost upon propagation of the strains.

Loading

Article metrics loading...

/content/journal/micro/10.1099/00221287-143-4-1211
1997-04-01
2024-12-14
Loading full text...

Full text loading...

/deliver/fulltext/micro/143/4/mic-143-4-1211.html?itemId=/content/journal/micro/10.1099/00221287-143-4-1211&mimeType=html&fmt=ahah

References

  1. Arst H. N. Jr, Sheerin A. 1996; Translational initiation competence,‘ leaky scanning’ and translational reinitiation in areA of Aspergillus nidulans. . Mol Microbiol 19:1019–1024
    [Google Scholar]
  2. Beushausen S., Bergold P., Sturner S., Elste A., Roytenberg V., Schwartz J. H., Bayley H. 1988; Two catalytic subunits of cAMP-dependent protein kinase generated by alternative splicing are expressed in Aplysia neurons.. Neuron 1:853–864
    [Google Scholar]
  3. Boorstein W. R., Craig E.A. 1990; Regulation of a yeast HSP70 gene by a cAMP responsive transcriptional control element.. EMBO J 9:2543–2553
    [Google Scholar]
  4. Calzone F. J., Britten R. J., Davison E. H. 1987; Mapping of gene transcription by nuclease protection assays and cDNA primer extension.. Methods Enzymol 152:611–632
    [Google Scholar]
  5. Døskeland S. O., Maronde E., Gjersten B. T. 1993; The genetic subtypes of cAMP-dependent protein kinase - functionally different or redundant ?. Biochim Biophys Acta 1178:249–258
    [Google Scholar]
  6. Goosen T., Bloemheuvel G., Gysler C., de Bie D.A., van den Broek H. W. J., Swart K. 1987; Transformation of Aspergillus niger using the homologous orotidine-5’-phosphate-decarboxylase gene.. Curr Genet 11:499–503
    [Google Scholar]
  7. Gounalaki N., Thireos G. 1994; Yaplp, a yeast transcriptional activator that mediates multidrug resistance, regulates the metabolic stress response.. EMBO J 13:4036–4041
    [Google Scholar]
  8. de Graaff L., van den Broeck H., Visser J. 1988; Isolation and expression of the Aspergillus nidulans pyruvate kinase gene.. Curr Genet 13:315–321
    [Google Scholar]
  9. Gross R.E., Bagchi S.Lu X., Rubin C.S. 1990; Cloning and characterisation and expression of the gene for the catalytic subunit of the cAMP-dependent protein kinase in Caenorhabditis elegans.. J Biol Chem 265:6896–6907
    [Google Scholar]
  10. Harmsen J. A. M., Kusters-van Someren M. A., Visser J. 1990; Cloning and expression of a second Aspergillus niger pectin lyase gene (pelA): indications of a pectin lyase gene family in Aspergillus niger.. Curr Genet 18:161–166
    [Google Scholar]
  11. Kobayashi N., McEntee K. 1993; Identification of cis and transcomponents of a novel heat shock stress regulatory pathway in Saccharomyces cerevisiae.. Mol Cell Biol 13:248–256
    [Google Scholar]
  12. 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 Aspergillus niger and Aspergillus nidulans.. Curr Genet 20:293–299
    [Google Scholar]
  13. Lalli E., Sassone-Corsi P. 1994; Signal transduction and gene regulation: the nuclear response to cAMP.. J Biol Chem 269:17359–17362
    [Google Scholar]
  14. Legiša M., Bencina M. 1994; Evidence for the activation of 6- phosphofructo-1-kinase by cAMP-dependent protein kinase in Aspergillus niger.. FEMS Microbiol Lett 118:327–334
    [Google Scholar]
  15. Legiša M., Grapulin-Gradisnik M. 1995; Sudden substrate dilution induces a higher rate of citric acid production by Aspergillus niger.. Appl Environ Microbiol 61:2732–2737
    [Google Scholar]
  16. Maeda T., Watanabe Y., Kunitomo H., Yamamoto M. 1994; Cloning of the pkal gene encoding the catalytic subunit of the cAMP-dependent protein kinase in Schizosaccharomyces pombe.. J Biol Chem 269:9632–9637
    [Google Scholar]
  17. Mann S. K. O., Firtel R. A. 1991; A developmentally regulated putative serine/threonine protein kinase is essential for development in Dictyostelium.. Mech Dev 35:89–101
    [Google Scholar]
  18. Mann S. K. O., Firtel R. A. 1993; cAMP-dependent protein kinase differentially regulates prestalk and prespore differentiation during Dictyostelium development.. Development 119:135–146
    [Google Scholar]
  19. Marchler G., Schuller C., Adam G., Ruis H. 1993; A Saccharomyces cerevisiae UAS element controlled by protein kinase A activates transcription in response to a variety of stress conditions.. EMBO J 12:1997–2003
    [Google Scholar]
  20. Mazon M. J., Behrens M. M., Morgado E., Portillo F. 1993; Low activity of the yeast cAMP-dependent protein kinase catalytic subunit Tpk3 is due to the poor expression of the TPK3 gene.. Eur J Biochem 213:501–506
    [Google Scholar]
  21. 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 Cell 7:1869–1878
    [Google Scholar]
  22. 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 USA 84:6–10
    [Google Scholar]
  23. de Oliveira J. C. F, Borges A. C. C., Marques M. V., Gomes S. L. 1994; Cloning and characterisation of the gene for the catalytic subunit of cAMP-dependent protein kinase in the aquatic fungus Blastocladiella emersonii.. Eur J Biochem 219:555–562
    [Google Scholar]
  24. Pastori R., Moreno S., Passeron S. 1985; Polymeric structure of the cyclic AMP-dependent protein kinase from the dimorphic fungus Mucor rouxii and purification of its catalytic subunit.. Mol Cell Biochem 69:55–66
    [Google Scholar]
  25. Pontecorvo G., Roper J. A., Hemmons L. J., MacDonald K.D., Bufton A. W. J. 1953; The genetics of Aspergillus nidulans.. Adv Genet 5:141–238
    [Google Scholar]
  26. Powers P. A., Pall M. L. 1980; Cyclic AMP-dependent protein kinase of Neurospora crassa.. Biochem Biophys Res Commun 95:701–706
    [Google Scholar]
  27. Rossi S., Moreno S. 1994; Regulation of protein kinase A subunits during germination of Mucor rouxii sporangiospores.. Eur J Biochem 222:501–506
    [Google Scholar]
  28. 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]
  29. Sanger F., Nicklen S., Coulson A. R. 1977; DNA sequencing with chain-terminating inhibitors.. Proc Natl Acad Sci USA 74:5463–5467
    [Google Scholar]
  30. Simoni M. N., Pelegrini O., Veron M., Kay R. R. 1992; Mutation of protein kinase A causes heterochronic development of Dictyostelium.. Nature 356:171–172
    [Google Scholar]
  31. Struhl K. 1985; Naturally occurring poly(dA-dT) sequences are upstream promoter elements for constitutive transcription in yeast.. Proc Natl Acad Sci USA 82:8419–8423
    [Google Scholar]
  32. 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 Biol 8:429–462
    [Google Scholar]
  33. Thevelein J. M. 1994; Signal transduction in yeast.. Yeast 10:1753–1790
    [Google Scholar]
  34. Toda T., Cameron S., Sass P., Zoller M., Wigler M. 1987a; Three different genes in S.cerevisiae encode the catalytic subunit of the cAMP-dependent protein kinase.. Cell 50:277–287
    [Google Scholar]
  35. Toda T., Cameron S., Sass P., Zoller M., Scott J.D., McMullen B., Hurwitz M., Krebs E.G., Wigler M. 1987b; Cloning and characterisation of BCY1, a locus encoding a regulatory subunit of the cyclic AMP-dependent protein kinase in Saccharomyces cerevisiae.. Mol Cell Biol 7:1371–1377
    [Google Scholar]
  36. Unkles S. E. 1992; Gene organisation in industrial filamentous fungi.In Applied Molecular Genetics of Filamentous Fungi, pp. Edited by J. R. Kinghorn & G. Turner. London: Blackie.. 28–53
    [Google Scholar]
  37. Visniac W., Santer M. 1957; The thiobacilli.. Bacteriol Rev 21:195–213
    [Google Scholar]
  38. Walsh D.A., van Patten S.M. 1994; Multiple pathway signal transduction by the cAMP-dependent protein kinase.. FASEB (Fed Am Soc Exp Biol) J 8:1227–1236
    [Google Scholar]
  39. wiemann S., Kinzel V., Pyerin W. 1992; Cloning of the Cα catalytic subunit of the bovine cAMP-dependent protein kinase.. Biochim Biophys Acta 1171:93–96
    [Google Scholar]
  40. Witteveen C. B. F., van de Vondervoort P. J. I., van den Broeck H. C., van Engelenburg F. A. C., de Graaff L. H., Hillebrand M. H. B. C., Schaap P. J., Visser J. 1993; Induction of glucose oxidase, catalase and lactonase in Aspergillus niger.. Curr Genet 24:408–416
    [Google Scholar]
/content/journal/micro/10.1099/00221287-143-4-1211
Loading
/content/journal/micro/10.1099/00221287-143-4-1211
Loading

Data & Media loading...

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