Artículo

Estamos trabajando para incorporar este artículo al repositorio
Consulte el artículo en la página del editor
Consulte la política de Acceso Abierto del editor

Abstract:

The protein kinase A (PKA) signaling pathway plays a role in regulating growth and differentiation in the dimorphic fungus Mucor circinelloides. PKA holoenzyme is comprised of two catalytic (C) and two regulatory (R) subunits. In M. circinelloides, four genes encode the PKAR1, PKAR2, PKAR3, and PKAR4 isoforms of R subunits. We have constructed null mutants and demonstrate that each isoform has a different role in growth and differentiation. The most striking finding is that pkaR4 is an essential gene, because only heterokaryons were obtained in knockout experiments. Heterokaryons with low levels of wild-type nuclei showed an impediment in the emission of the germ tube, suggesting a pivotal role of this gene in germ tube emergence. The remaining null strains showed different alterations in germ tube emergence, sporulation, and volume of the mother cell. The pkaR2 null mutant showed an accelerated germ tube emission and was the only mutant that germinated under anaerobic conditions when glycine was used as a nitrogen source, suggesting that pkaR2 participates in germ tube emergence by repressing it. From the measurement of the mRNA and protein levels of each isoform in the wild-type and knockout strains, it can be concluded that the expression of each subunit has its own mechanism of differential regulation. The PKAR1 and PKAR2 isoforms are posttranslationally modified by ubiquitylation, suggesting another regulation point in the specificity of the signal transduction. The results indicate that each R isoform has a different role in M. circinelloides physiology, controlling the dimorphism and contributing to the specificity of cyclic AMP (cAMP)-PKA pathway. © 2012, American Society for Microbiology. All Rights Reserved.

Registro:

Documento: Artículo
Título:Protein kinase A regulatory subunit isoforms regulate growth and differentiation in Mucor circinelloides: Essential role of pkaR4
Autor:Ocampo, J.; McCormack, B.; Navarro, E.; Moreno, S.; Garre, V.; Rossi, S.
Filiación:Departamento de Química Biológica, Facultad de Ciencias Exactas y Naturales, Universidad de Buenos Aires, Ciudad Universitaria, Buenos Aires, Argentina
Departamento de Genética y Microbiología (Unidad Asociada al IQFR-CSIC), Facultad de Biología, Universidad de Murcia, Murcia, Spain
Palabras clave:cyclic AMP dependent protein kinase; fungal protein; holoenzyme; isoprotein; article; enzymology; fungus spore; gene deletion; genetics; growth, development and aging; metabolism; morphogenesis; Mucor; protein subunit; signal transduction; ubiquitination; Cyclic AMP-Dependent Protein Kinases; Fungal Proteins; Gene Deletion; Holoenzymes; Morphogenesis; Mucor; Protein Isoforms; Protein Subunits; Signal Transduction; Spores, Fungal; Ubiquitination; Fungi; Mucor circinelloides
Año:2012
Volumen:11
Número:8
Página de inicio:989
Página de fin:1002
DOI: http://dx.doi.org/10.1128/EC.00017-12
Título revista:Eukaryotic Cell
Título revista abreviado:Eukaryotic Cell
ISSN:15359778
CODEN:ECUEA
CAS:Cyclic AMP-Dependent Protein Kinases, 2.7.11.11; Fungal Proteins; Holoenzymes; Protein Isoforms; Protein Subunits
Registro:https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_15359778_v11_n8_p989_Ocampo

Referencias:

  • Adachi, K., Hamer, J.E., Divergent cAMP signaling pathways regulate growth and pathogenesis in the rice blast fungus Magnaporthe grisea (1998) Plant Cell, 10, pp. 1361-1373
  • Banno, S., A catalytic subunit of cyclic AMP dependent protein kinase, PKAC-1, regulates asexual differentiation in Neurospora crassa (2005) Genes Genet. Syst, 80, pp. 25-34
  • Bartnicki-García, S., Nickerson, W.J., Nutrition, growth and morphogenesis of Mucor rouxii (1962) J. Bacteriol, 84, pp. 841-858
  • Biswas, S., van Dijck, P., Datta, A., Environmental sensing and signal transduction pathways regulating morphopathogenic determinants of Candida albicans (2007) Microbiol Mol. Biol. Rev, 71, pp. 348-376
  • Borges-Walmsley, M.I., Walmsley, A.R., cAMP signalling in pathogenic fungi: Control of dimorphic switching and pathogenicity (2000) Trends Microbiol, 8, pp. 133-141
  • Brown, J., Zygomycosis: An emerging fungal infection (2005) Am. J. Health Syst. Pharm, 62, pp. 2593-2596
  • Bruno, K.S., Aramayo, R., Minke, P.F., Metzenber, R.L., Plamann, M., Loss of growth polarity and mislocalization of septa in a Neurospora mutant altered in the regulatory subunit of cAMP-dependent protein kinase (1996) EMBO J, 15, pp. 5772-5782
  • Cassola, A., Candida albicans lacking the gene encoding the regulatory subunit of protein kinase A displays a defect in hyphal formation and an altered localization of the catalytic subunit (2004) Eukaryot. Cell, 3, pp. 190-199
  • Cervantes-Chávez, J.A., Ruiz-Herrera, J., The regulatory subunit of protein kinase A promotes hyphal growth and plays an essential role in Yarrowia lipolytica (2007) FEMS Yeast Res, 7, pp. 929-940
  • Chayakulkeeree, M., Ghannoum, M.A., Perfect, J.R., Zygomycosis: The re-emerging fungal infection (2006) Eur. J. Clin. Microbiol. Infect. Dis, 25, pp. 215-229
  • D'Souza, C.A., Cyclic AMP-dependent protein kinase controls virulence of the fungal pathogen Cryptococcus neoformans (2001) Mol. Cell. Biol, 21, pp. 3179-3191
  • Durrenberger, F., Wong, K., Kronstad, J.W., Identification of a cAMP-dependent protein kinase catalytic subunit required for virulence and morphogenesis in Ustilago maydis (1998) Proc. Natl. Acad. Sci. U. S. A, 95, pp. 5684-5689
  • Gilman, A., Presentation of the Academy Medal to Harry Eagle (1970) M. D. Bull. N. Y. Acad. Med, 46, pp. 666-669
  • Gold, S.E., Duncan, G., Barrett, K., Kronstad, J., cAMP regulates morphogenesis in the fungal pathogen Ustilago maydis (1994) Genes Dev, 8, pp. 2805-2816
  • Gutierrez, A., Lopez-Garcia, S., Garre, V., High reliability transformation of the basal fungus Mucor circinelloides by electroporation (2011) J. Microbiol. Methods, 84, pp. 442-446
  • Harris, S.D., Cell polarity in filamentous fungi: Shaping the mold (2006) Int. Rev. Cytol, 251, pp. 41-77
  • He, X., Zhang, J., Higher duplicability of less important genes in yeast genomes (2006) Mol. Biol. Evol, 23, pp. 144-151
  • Ichinomiya, M., Uchida, H., Koshi, Y., Ohta, A., Horiuchi, H., A protein kinase C-encoding gene, pkcA, is essential to the viability of the filamentous fungus Aspergillus nidulans (2007) Biosci. Biotechnol. Biochem, 71, pp. 2787-2799
  • Jarnaess, E., Tasken, K., Spatiotemporal control of cAMP signaling processes by anchored signalling complexes (2007) Biochem. Soc. Trans, 35, pp. 931-937
  • Kinderman, F.S., A dynamic mechanism for AKAP binding to RII isoforms of cAMP-dependent protein kinase (2006) Mol. Cell, 24, pp. 397-408
  • Klein, B.S., Tebbets, B., Dimorphism and virulence in fungi (2007) Curr. Opin. Microbiol, 10, pp. 314-319
  • Kontoyiannis, D.P., Zygomycosis in a tertiary-care cancer center in the era of Aspergillus-active antifungal therapy: A case-control observational study of 27 recent cases (2005) J. Infect. Dis, 191, pp. 1350-1360
  • Li, C.H., Sporangiospore size dimorphism is linked to virulence of Mucor circinelloides (2011) PLoS Pathog, 7, p. 1002086
  • Lu, Z., Hunter, T., Degradation of activated protein kinases by ubiquitination (2009) Annu. Review Biochem, 78, pp. 435-475
  • Lübbehusen, T., Protein kinase A is involved in the control of morphology and branching during aerobic growth of Mucor circinelloides (2004) Microbiology, 150, pp. 143-150
  • Marques, M.V., Gomes, L.S., Cloning and structural analysis of the gene for the regulatory subunit of cAMP-dependent protein kinase in Blastocladiella emersonii (1992) J. Biol. Chem, 267, pp. 17201-17207
  • Marr, K.A., Carter, R.A., Crippa, F., Wald, A., Corey, L., Epidemiology and outcome of mould infections in hematopoietic stem cell transplant recipients (2002) Clin. Infect. Dis, 34, pp. 909-917
  • Navarro, E., A negative regulator of light-inducible carotenogenesis in Mucor circinelloides (2001) Mol. Genet. Genomics, 266, pp. 463-470
  • Nemecek, J.C., Wuthrich, M., Klein, B.S., Global control of dimorphism and virulence in fungi (2006) Science, 312, pp. 583-588
  • Nicolas, F.E., de Haro, J.P., Torres-Martinez, S., Ruiz-Vazquez, R.M., Mutants defective in a Mucor circinelloides dicer-like gene are not compromised in siRNA silencing but display developmental defects (2007) Fungal Genet. Biol, 44, pp. 504-516
  • Ocampo, J., A subunit of protein kinase A regulates growth and differentiation in the fungus Mucor circinelloides (2009) Eukaryot Cell, 8, pp. 933-944
  • Ocampo, J., Moreno, S., Rossi, S., PKA from Mucor circinelloides: Model to study the role of linker I in the interaction between R and C subunits (2007) Biochem. Biophys. Res. Commun, 362, pp. 721-726
  • Orlowski, M., Mucor dimorphism (1991) Microbiol. Rev, 55, pp. 234-258
  • Osmani, A.H., Oakley, B.R., Osmani, S.A., Identification and analysis of essential Aspergillus nidulans genes using the heterokaryon rescue technique (2006) Nat. Protoc, 1, pp. 2517-2526
  • Pereyra, E., Mizyrycki, C., Moreno, S., Threshold level of protein kinase A activity and polarized growth in Mucor rouxii (2000) Microbiology, 146, pp. 1949-1958
  • Pereyra, E., Zaremberg, V., Moreno, S., Effect of dibutyryl cAMP on growth and morphology of germinating Mucor rouxii sporangiospores (1992) Exp. Mycol, 16, pp. 93-101
  • Quiles-Rosillo, M.D., Ruiz-Vazquez, R.M., Torres-Martinez, S., Garre, V., Cloning, characterization and heterologous expression of the Blakesleatrispora gene encoding orotidine-5=-monophosphate decarboxylase (2003) FEMS Microbiol. Lett, 222, pp. 229-236
  • Ribes, J.A., Vanover-Sams, C.L., Baker, D.J., Zygomycetes in human disease (2000) Clin. Microbiol. Rev, 13, pp. 236-301
  • Rinaldi, J., Ocampo, J., Rossi, S., Moreno, S., A novel activating effect of the regulatory subunit of protein kinase A on catalytic subunit activity (2008) Arch. Biochem. Biophys, 480, pp. 95-103
  • Roden, M.M., Epidemiology and outcome of zygomycosis: A review of 929 reported cases (2005) Clin. Infect. Dis, 41, pp. 634-653
  • Roncero, M.I., Zabala, C., Cerda-Olmedo, E., Mutagenesis in multinucleate cells: The effects of N-methyl-N=-nitro-N-nitrosoguanidine on Phycomyces spores (1984) Mutat. Res, 125, pp. 195-204
  • Roskoski, R., Assays of protein kinase (1983) Methods Enzymol, 99, pp. 3-6
  • Ruiz-Perez, V.L., Murillo, F.J., Torres-Martinez, S., PkpA, a novel Phycomyces blakesleeanus serine/threonine protein kinase (1995) Curr. Genet, 28, pp. 309-316
  • Sambrook, J., Russell, D.W., (2001) Molecular Cloning: A Laboratory Manual, , 3rd ed. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY
  • Sarma, G.N., Structure of D-AKAP2:PKA RI complex: Insights into AKAP specificity and selectivity (2010) Structure, 18, pp. 155-166
  • Saudohar, M., Cyclic AMP-dependent protein kinase is involved in morphogenesis in Aspergillus niger (2002) Microbiology, 148, pp. 2635-2645
  • Schipper, M.A., Induced azygospore formation in Mucor (Rhizomucor) pusillus by Absidia corymbifera (1976) Antonie Van Leeuwenhoek, 42, pp. 141-144
  • Silva, F., A RING-finger protein regulates carotenogenesis via proteolysis-independent ubiquitylation of a white collar-1-like activator (2008) Mol. Microbiol, 70, pp. 1026-1036
  • Spellberg, B., Edwards Jr., J., Ibrahim, A., Novel perspectives on mucormycosis: Pathophysiology, presentation, and management (2005) Clin. Microbiol. Rev, 18, pp. 556-569
  • Stiefel, J., Suppressors of an adenylate cyclase deletion in the fission yeast Schizosaccharomyces pombe (2004) Eukaryot. Cell, 3, pp. 610-619
  • Takano, Y., Komeda, K., Kojima, K., Okuno, T., Proper regulation of cyclic AMP-dependent protein kinase is required for growth, conidiation, and appressorium function in the anthracnose fungus Colletotrichum lagenarium (2001) Mol. Plant Microbe Interact, 14, pp. 1149-1157
  • Toda, T., Cloning and characterization of BCY1, a locus encoding a regulatory subunit of the cyclic AMP-dependent protein kinase in Saccharomyces cerevisiae (1987) Mol. Cell. Biol, 7, pp. 1371-1377
  • Wolff, A.M., Appel, K.F., Petersen, J.B., Poulsen, U., Arnau, J., Identification and analysis of genes involved in the control of dimorphism in Mucor circinelloides (syn. racemosus) (2002) FEMS Yeast. Res, 2, pp. 203-213
  • Zhao, W., Deletion of the regulatory subunit of protein kinase A in Aspergillus fumigatus alters morphology, sensitivity to oxidative damage, and virulence (2006) Infect. Immun, 74, pp. 4865-4874
  • Ziv, C., Gorovits, R., Yarden, O., Carbon source affects PKA-dependent polarity of Neurospora crassa in a CRE-1-dependent and independent manner (2008) Fungal Genet. Biol, 45, pp. 103-116

Citas:

---------- APA ----------
Ocampo, J., McCormack, B., Navarro, E., Moreno, S., Garre, V. & Rossi, S. (2012) . Protein kinase A regulatory subunit isoforms regulate growth and differentiation in Mucor circinelloides: Essential role of pkaR4. Eukaryotic Cell, 11(8), 989-1002.
http://dx.doi.org/10.1128/EC.00017-12
---------- CHICAGO ----------
Ocampo, J., McCormack, B., Navarro, E., Moreno, S., Garre, V., Rossi, S. "Protein kinase A regulatory subunit isoforms regulate growth and differentiation in Mucor circinelloides: Essential role of pkaR4" . Eukaryotic Cell 11, no. 8 (2012) : 989-1002.
http://dx.doi.org/10.1128/EC.00017-12
---------- MLA ----------
Ocampo, J., McCormack, B., Navarro, E., Moreno, S., Garre, V., Rossi, S. "Protein kinase A regulatory subunit isoforms regulate growth and differentiation in Mucor circinelloides: Essential role of pkaR4" . Eukaryotic Cell, vol. 11, no. 8, 2012, pp. 989-1002.
http://dx.doi.org/10.1128/EC.00017-12
---------- VANCOUVER ----------
Ocampo, J., McCormack, B., Navarro, E., Moreno, S., Garre, V., Rossi, S. Protein kinase A regulatory subunit isoforms regulate growth and differentiation in Mucor circinelloides: Essential role of pkaR4. Eukaryotic Cell. 2012;11(8):989-1002.
http://dx.doi.org/10.1128/EC.00017-12