1887

Abstract

A model system to study the involvement of cAMP-mediated regulation of a cellular process such as hyphal morphogenesis was investigated. Impairment of polarized growth was observed when sporangiospores were grown in the presence of -cAMP analogues and of SQ 65,442, a cAMP phosphodiesterase inhibitor. Together with an effect on isodiametric growth, there was increased pigmentation, increased cell fragility and loss of cell adhesiveness. The total effect on morphology was attained even after adding the compounds shortly before germ-tube emergence; when added after this time growth continued in a non-polarized form and rounding of the germ tip was observed. The morphological effect was observed under all the nutritional and environmental conditions studied (aerobic conditions and defined medium with maltose or glucose, Casamino acids medium with glucose, or rich medium; anaerobic conditions with rich medium; and following a shift from anaerobiosis to aerobiosis). The time of germ-tube emergence, and the size of the cell at this time, was dependent on the growth medium. Protein kinase A (PKA) specific activity was followed during the germination process under three growth conditions. It was found that the total activity of PKA correlated with differentiation and not with growth, and that the total specific activity at the time of germination was the same, independent of the culture medium. The time of germ-tube emergence correlated with the time of attainment of a threshold level of PKA total specific activity. The concentration of dibutyryl-cAMP needed to promote isodiametric growth correlated with the total units of activity of PKA to be activated per cell. It was concluded that PKA is involved in the morphogenetic process of the fungus grown under all the nutritional and ambient conditions tested.

Loading

Article metrics loading...

/content/journal/micro/10.1099/00221287-146-8-1949
2000-08-01
2020-03-28
Loading full text...

Full text loading...

/deliver/fulltext/micro/146/8/1461949a.html?itemId=/content/journal/micro/10.1099/00221287-146-8-1949&mimeType=html&fmt=ahah

References

  1. Alspaugh J. A., Perfect J. R., Heitman J.. 1998; Signal transduction pathways regulating differentiation and pathogenicity of Cryptococcus neoformans. Fungal Genet Biol25:1–14[CrossRef]
    [Google Scholar]
  2. Bartnicki-Garcı́a S.. 1990; Role of vesicles in apical growth and a new mathematical model of hyphal morphogenesis. In Tip Growth in Plant and Fungal Cells pp.211–232Edited by Heath I. B. . San Diego: Academic Press;
    [Google Scholar]
  3. Bartnicki-Garcı́a S.. 1996; The hypha: unifying thread of the fungal kingdom. In A Century of Mycology pp.105–133Edited by Sutton B. C.. Cambridge: Cambridge University Press;
    [Google Scholar]
  4. Bartnicki-Garcı́a S., Nickerson W. J.. 1962; Nutrition, growth and morphogenesis of Mucor rouxii. J Bacteriol83:841–858
    [Google Scholar]
  5. Bradford M.. 1976; A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein–dye binding. Anal Biochem72:248–254[CrossRef]
    [Google Scholar]
  6. Braunmann T., Erneux C., Petridis G., Stohrer W., Jastorff B.. 1986; Hydrolysis of cyclic nucleotides by a purified cGMP-phosphodiesterase; structural requirements for hydrolysis. Biochim Biophys Acta871:199–206[CrossRef]
    [Google Scholar]
  7. 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]
  8. Dandekar S., Modi V. V.. 1980; Involvement of cyclic AMP in carotenogenesis and cell differentiation in Blakeslea trispora. Biochim Biophys Acta628:398–406[CrossRef]
    [Google Scholar]
  9. Doerner P., Jorgensen J., You R., Steppuhn J., Lamb C.. 1996; Control of root growth and development by cyclin expression. Nature380:520–523[CrossRef]
    [Google Scholar]
  10. Edgar B. A., O’Farrell P. H.. 1989; Genetic control of cell division patterns in the Drosophila embryo. Cell57:177–187[CrossRef]
    [Google Scholar]
  11. Glass W. F. Jr, Kreisberg J. I.. 1993; Regulation of integrin-mediated adhesion at focal contacts by cyclic AMP. J Cell Physiol157:296–306[CrossRef]
    [Google Scholar]
  12. Guthmann M., Pastori R., Moreno S.. 1990; Polyamines and basic proteins stimulate activation by cAMP and catalytic activity of Mucor rouxii cAMP dependent protein kinase. Cell Signal2:395–402[CrossRef]
    [Google Scholar]
  13. Haidle C. W., Storck R.. 1966; Control of dimorphism in Mucor rouxii. J Bacteriol92:1236–1244
    [Google Scholar]
  14. Hamer J. E., Talbot N. J.. 1998; Infection-related development in the rice blast fungus Magnaporthe grisea. Curr Opin Microbiol1:693–697[CrossRef]
    [Google Scholar]
  15. Heath I. B.. 1994; The cytoskeleton in hyphal growth, organelle movements and mitosis. In The Mycota: a Comprehensive Treatise on Fungi as Experimental Systems for Basic and Applied Researchvol. 1Growth, Differentiation and Sexuality pp.43–65Edited by Wessels J. H. G., Meinhardt F.. Berlin: Springer;
    [Google Scholar]
  16. Hemerly A., Engler J., Bergounioux C., Montagu M., Engler G., Inze D., Ferrerira P.. 1995; Dominant negative mutants of the Cdc2 kinase uncouple cell division from iterative plant development. EMBO J14:3925–3936
    [Google Scholar]
  17. Hohl N., Galland P., Senger H.. 1992; Altered pterin patterns in photobehavioral mutants of Phycomyces blakesleeanus. Photochem Photobiol55:239–245[CrossRef]
    [Google Scholar]
  18. Jackson S. L., Heath I. B.. 1993; Roles of calcium ions in hyphal tip growth. Microbiol Rev57:367–382
    [Google Scholar]
  19. Jones B. E., Bu’Lock J. D.. 1977; The effect of N6,O2′-dibutyryl adenosine-3′,5′-cyclic monophosphate on morphogenesis in mucorales. J Gen Microbiol103:29–36[CrossRef]
    [Google Scholar]
  20. Kronstad J. W.. 1997; Virulence and cAMP in smuts, blasts and blights. Trends Plant Sci2:193–200[CrossRef]
    [Google Scholar]
  21. Kronstad J., De Maria A., Funnell D., Laidlaw R. D., Lee N., Moniz de Sá M., Ramesh M.. 1998; Signalling via cAMP in fungi: interconnections with mitogen-activated protein kinase pathways. Arch Microbiol170:395–404[CrossRef]
    [Google Scholar]
  22. Larsen A. D., Sypherd P.. 1974; Cyclic adenosine 3′,5′-monophosphate and morphogenesis in Mucor racemosus. J Bacteriol117:432–438
    [Google Scholar]
  23. Madhani H. D., Fink G. R.. 1998; The control of filamentous differentiation and virulence in fungi. Trends Cell Biol8:348–353[CrossRef]
    [Google Scholar]
  24. Money N. P.. 1994; Osmotic adjustment and the role of turgor in mycelial fungi. In The Mycota: a Comprehensive Treatise on Fungi as Experimental Systems for Basic and Applied Researchvol. 1Growth, Differentiation and Sexuality pp.67–88Edited by Wessels J. H. G., Meinhardt F.. Berlin: Springer;
    [Google Scholar]
  25. Moreno S., Passeron S.. 1980; Further studies on cAMP-dependent protein kinase from the dimorphic fungus Mucor rouxii. Arch Biochem Biophys199:321–330[CrossRef]
    [Google Scholar]
  26. Moreno S., Pastori R., Passeron S.. 1983; Protein kinase of Mucor rouxii: unshielding of new cyclic AMP binding sites upon dissociation of the ternary complex holoenzyme–cyclic AMP. Mol Cell Biochem52:13–16
    [Google Scholar]
  27. Øgreid D., Ekanger R., Suva R., Miller J., Sturm P., Corbin J. D., Døskeland S. O.. 1985; Activation of protein kinase isozymes by cyclic nucleotide analogues used singly or in combination. Eur J Biochem150:219–227[CrossRef]
    [Google Scholar]
  28. Orlowski M.. 1991; Mucor dimorphism. Microbiol Rev55:234–258
    [Google Scholar]
  29. Orlowski M.. 1995; Gene expression in Mucor dimorphism. Can J Bot73:S326–S334[CrossRef]
    [Google Scholar]
  30. Paveto C., Passeron S., Corbin J. D., Moreno S.. 1989; Two different intrachain cAMP sites in the cAMP-dependent protein kinase from the dimorphic fungus Mucor rouxii. Eur J Biochem179:429–434[CrossRef]
    [Google Scholar]
  31. Pereyra E., Zaremberg V., Moreno S.. 1992; Effect of dibutyryl-cAMP on growth and morphology of germinating Mucor rouxii sporangiospores. Exp Mycol16:93–101[CrossRef]
    [Google Scholar]
  32. Roskoski R.. 1983; Assay of protein kinase. Methods Enzymol99:3–6
    [Google Scholar]
  33. Rossi S., Moreno S.. 1994; Regulation of protein kinase A subunits during germination of Mucor rouxii sporangiospores. Eur J Biochem222:501–506[CrossRef]
    [Google Scholar]
  34. Roze L. V., Mahanti N., Mehigh R., McConnell D. G., Linz J. E.. 1999; Evidence that MRas1 and MRas3 proteins are associated with distinct cellular functions during growth and morphogenesis in the fungus Mucor racemosus. Fungal Genet Biol28:171–180[CrossRef]
    [Google Scholar]
  35. Sabanero M.. 1994; Effects of cytochalasins on growth of Mucor rouxii . Microbios80:91–97
    [Google Scholar]
  36. Sietsma J. H., Wessels J. H. G.. 1994; Apical wall biogenesis. In The Mycotavol. 1 pp.126–141Edited by Wessels J. H. G., Meinhardt F.. Berlin: Springer;
    [Google Scholar]
  37. Tomes C., Moreno S.. 1990; cAMP analogs and selective inhibitors used to study low K m Mucor rouxii cAMP phosphodiesterase. Int J Biochem22:1047–1051[CrossRef]
    [Google Scholar]
  38. Tomes C., Rossi S., Moreno S.. 1993; Isobutyl-methyl-xanthine and other classical cyclic nucleotide phosphodiesterase inhibitors affect cAMP-dependent protein kinase activity. Cell Signal5:615–621[CrossRef]
    [Google Scholar]
  39. Van Haastert P. J. M., Dijkgraaf P. A. M., Konijn T. M., Garcia Abbad E., Petridis G., Jastorff B.. 1983; Substrate specificity of cyclic nucleotide phosphodiesterase from beef heart and from Dictyostelium discoideum. Eur J Biochem131:659–666[CrossRef]
    [Google Scholar]
  40. Van Lookeren-Campagne M. M., Villalba Dı́az F., Jastorff B., Winkler E., Genieser H., Kessin R. H.. 1990; Characterization of the yeast low K m phosphodiesterase with cAMP analogues: applications in mammalian cells that express the yeast PDE2 gene. J Biol Chem265:5847–5854
    [Google Scholar]
  41. Yang S., Dickman M. B.. 1999; Colletotrichum trifolii mutants disrupted in the catalytic subunit of cAMP-dependent protein kinase are nonpathogenic. Mol Plant–Microbe Interact12:430–439[CrossRef]
    [Google Scholar]
  42. Ye X. S., Lee S., Wolkow T. D., McGuire S. L., Hamer J. E., Wood C. G., Osmani S. A.. 1999; Interaction between developmental and cell cycle regulators is required for morphogenesis in Aspergillus nidulans. EMBO J18:6994–7001[CrossRef]
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/micro/10.1099/00221287-146-8-1949
Loading
/content/journal/micro/10.1099/00221287-146-8-1949
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