G-protein and cAMP-mediated signaling in aspergilli: a genomic perspective.

We have carried out an in silico exploration of the genomes of Aspergillus nidulans, Aspergillus fumigatus, and Aspergillus oryzae, and identified components of G-protein/cAMP-mediated signaling. Putative G-protein coupled receptors (GPCRs) were distributed over nine classes. The GPCRs within classes were well conserved among aspergilli but varied in other ascomycetes. As previously observed in A. nidulans and other fungi, three Galpha, one Gbeta, and one Ggamma subunits of G proteins were identified in A. fumigatus, whereas an additional likely non-functional Galpha subunit was present in A. oryzae. While most fungal species had five proteins containing the regulator of G-protein signaling (RGS) domain predicted to participate in attenuation of G-protein signaling, A. fumigatus and A. oryzae had an additional RGS protein (RgsD) related to RgsA of A. nidulans. Genes encoding adenylate cyclase, a regulatory subunit and two catalytic subunits of the cAMP-dependent protein kinase, were also identified in the three aspergilli. Finally, regulators of cAMP signaling including low- and high-affinity phosphodiesterases were identified. Taken together, our data indicate a striking diversity at the GPCR level, but little diversity of components at the G-protein and cAMP-signaling level. This may reflect the abilities of these fungi to adapt to various ecological niches and to integrate diverse environmental cues into highly conserved cellular processes.

[1]  William H. Majoros,et al.  Genomic sequence of the pathogenic and allergenic filamentous fungus Aspergillus fumigatus , 2005, Nature.

[2]  William H. Majoros,et al.  Corrigendum: Genomic sequence of the pathogenic and allergenic filamentous fungus Aspergillus fumigatus , 2006, Nature.

[3]  Jae-Hyuk Yu,et al.  Multiple Roles of a Heterotrimeric G-Protein γ-Subunit in Governing Growth and Development of Aspergillus nidulans , 2005, Genetics.

[4]  Christina A. Cuomo,et al.  Sequencing of Aspergillus nidulans and comparative analysis with A. fumigatus and A. oryzae , 2005, Nature.

[5]  Kap-Hoon Han,et al.  The gprA and gprB genes encode putative G protein‐coupled receptors required for self‐fertilization in Aspergillus nidulans , 2004, Molecular microbiology.

[6]  Alan M. Jones,et al.  A Seven-Transmembrane RGS Protein That Modulates Plant Cell Proliferation , 2003, Science.

[7]  S. Pöggeler Genomic evidence for mating abilities in the asexual pathogen Aspergillus fumigatus , 2002, Current Genetics.

[8]  J. Hirsch,et al.  GPR1 encodes a putative G protein‐coupled receptor that associates with the Gpa2p Gα subunit and functions in a Ras‐independent pathway , 1998, The EMBO journal.

[9]  M. Bölker Sex and crime: heterotrimeric G proteins in fungal mating and pathogenesis. , 1998, Fungal genetics and biology : FG & B.

[10]  P. Dyer,et al.  Genomics reveals sexual secrets of Aspergillus. , 2003, Microbiology.

[11]  M. Yaffe,et al.  Temperature-sensitive yeast mutants defective in mitochondrial inheritance , 1990, The Journal of cell biology.

[12]  Manuel G. Claros,et al.  TopPred II: an improved software for membrane protein structure predictions , 1994, Comput. Appl. Biosci..

[13]  J. Latgé The pathobiology of Aspergillus fumigatus. , 2001, Trends in microbiology.

[14]  J. Latgé,et al.  Evidence for Sexuality in the Opportunistic Fungal Pathogen Aspergillus fumigatus , 2005, Current Biology.

[15]  J. D. de Winde,et al.  A Saccharomyces cerevisiae G‐protein coupled receptor, Gpr1, is specifically required for glucose activation of the cAMP pathway during the transition to growth on glucose , 1999, Molecular microbiology.

[16]  Cathryn J. Rehmeyer,et al.  The genome sequence of the rice blast fungus Magnaporthe grisea , 2005, Nature.

[17]  Kay Hofmann,et al.  Tmbase-A database of membrane spanning protein segments , 1993 .

[18]  B. Jahn,et al.  cAMP signaling in Aspergillus fumigatus is involved in the regulation of the virulence gene pksP and in defense against killing by macrophages , 2003, Molecular Genetics and Genomics.

[19]  C. S. Hoffman,et al.  Glucose monitoring in fission yeast via the Gpa2 galpha, the git5 Gbeta and the git3 putative glucose receptor. , 2000, Genetics.

[20]  R. Dean,et al.  Novel G-protein-coupled receptor-like proteins in the plant pathogenic fungus Magnaporthe grisea , 2005, Genome Biology.

[21]  Olivier Poch,et al.  A comprehensive comparison of multiple sequence alignment programs , 1999, Nucleic Acids Res..

[22]  J. Chant,et al.  Rax1, a protein required for the establishment of the bipolar budding pattern in yeast. , 2004, Gene.

[23]  J. Davey,et al.  Schizosaccharomyces pombe map3+ encodes the putative M-factor receptor , 1993, Molecular and cellular biology.

[24]  G. May,et al.  A Mitogen-Activated Protein Kinase That Senses Nitrogen Regulates Conidial Germination and Growth in Aspergillus fumigatus , 2004, Eukaryotic Cell.

[25]  Jae-Hyuk Yu,et al.  The Aspergillus nidulans sfaD gene encodes a G protein β subunit that is required for normal growth and repression of sporulation , 1999, The EMBO journal.

[26]  P. Thomas,et al.  Identification, classification, and partial characterization of genes in humans and other vertebrates homologous to a fish membrane progestin receptor , 2003, Proceedings of the National Academy of Sciences of the United States of America.

[27]  G. Sprague,,et al.  Evidence the yeast STE3 gene encodes a receptor for the peptide pheromone a factor: gene sequence and implications for the structure of the presumed receptor. , 1986, Proceedings of the National Academy of Sciences of the United States of America.

[28]  Dong-Min Han,et al.  The GanB Galpha-protein negatively regulates asexual sporulation and plays a positive role in conidial germination in Aspergillus nidulans. , 2004, Genetics.

[29]  A. Krogh,et al.  Predicting transmembrane protein topology with a hidden Markov model: application to complete genomes. , 2001, Journal of molecular biology.

[30]  Nancy Y. Villa,et al.  Metalloregulation of yeast membrane steroid receptor homologs. , 2004, Proceedings of the National Academy of Sciences of the United States of America.

[31]  A. Brakhage,et al.  The Cyclic AMP-Dependent Protein Kinase A Network Regulates Development and Virulence in Aspergillus fumigatus , 2004, Infection and Immunity.

[32]  J. Heitman,et al.  Signal Transduction Cascades Regulating Fungal Development and Virulence , 2000, Microbiology and Molecular Biology Reviews.

[33]  K. Hoe,et al.  Isolation of a Novel Gene fromSchizosaccharomyces pombe: stm1+ Encoding a Seven-transmembrane Loop Protein That May Couple with the Heterotrimeric Gα2 Protein, Gpa2* , 2001, The Journal of Biological Chemistry.

[34]  Thomas L. Madden,et al.  Gapped BLAST and PSI-BLAST: a new generation of protein database search programs. , 1997, Nucleic acids research.

[35]  Kap-Hoon Han,et al.  A putative G protein‐coupled receptor negatively controls sexual development in Aspergillus nidulans , 2004, Molecular microbiology.

[36]  Michael Freitag,et al.  Lessons from the Genome Sequence of Neurospora crassa: Tracing the Path from Genomic Blueprint to Multicellular Organism , 2004, Microbiology and Molecular Biology Reviews.

[37]  R. Dean,et al.  An eight-cysteine-containing CFEM domain unique to a group of fungal membrane proteins. , 2003, Trends in biochemical sciences.

[38]  J. Sweigard,et al.  Magnaporthe grisea Pth11p Is a Novel Plasma Membrane Protein That Mediates Appressorium Differentiation in Response to Inductive Substrate Cues , 1999, Plant Cell.

[39]  B. N. Lee,et al.  FluG and flbA function interdependently to initiate conidiophore development in Aspergillus nidulans through brlA beta activation. , 1996, The EMBO journal.

[40]  Jae-Hyuk Yu,et al.  The Aspergillus FlbA RGS domain protein antagonizes G protein signaling to block proliferation and allow development. , 1996, The EMBO journal.

[41]  D. Hereld,et al.  A cAMP receptor-like G protein-coupled receptor with roles in growth regulation and development. , 2004, Developmental biology.

[42]  Christophe d'Enfert,et al.  cAMP and ras signalling independently control spore germination in the filamentous fungus Aspergillus nidulans , 2002, Molecular microbiology.

[43]  C. d’Enfert,et al.  The Heterotrimeric G-Protein GanB(α)-SfaD(β)-GpgA(γ) Is a Carbon Source Sensor Involved in Early cAMP-Dependent Germination in Aspergillus nidulans , 2005, Genetics.

[44]  Sudhir Kumar,et al.  MEGA2: molecular evolutionary genetics analysis software , 2001, Bioinform..

[45]  E. Mauceli,et al.  The genome sequence of the filamentous fungus Neurospora crassa , 2003, Nature.

[46]  J. Thorner,et al.  Regulation of G protein-initiated signal transduction in yeast: paradigms and principles. , 2001, Annual review of biochemistry.

[47]  K. Isono,et al.  Genome sequencing and analysis of Aspergillus oryzae , 2005, Nature.

[48]  Norman W. Paton,et al.  CADRE: the Central Aspergillus Data REpository. , 2004 .

[49]  A. Krogh,et al.  A combined transmembrane topology and signal peptide prediction method. , 2004, Journal of molecular biology.

[50]  Jae-Hyuk Yu,et al.  Regulators of G‐protein signalling in Aspergillus nidulans: RgsA downregulates stress response and stimulates asexual sporulation through attenuation of GanB (Gα) signalling , 2004, Molecular microbiology.

[51]  Leland H. Hartwell,et al.  The yeast α-factor receptor: structural properties deduced from the sequence of the STE2 gene , 1985 .

[52]  K. Kitamura,et al.  The Schizosaccharomyces pombe mam2 gene encodes a putative pheromone receptor which has a significant homology with the Saccharomyces cerevisiae Ste2 protein. , 1991, The EMBO journal.

[53]  J. Heitman,et al.  The Gα Protein Gpa2 Controls Yeast Differentiation by Interacting with Kelch Repeat Proteins that Mimic Gβ Subunits , 2002 .

[54]  Bee-Na Lee,et al.  Overexpression of fIbA, an early regulator of Aspergillus asexual sporulation, leads to activation of brIA and premature initiation of development , 1994, Molecular microbiology.

[55]  K. Shimizu,et al.  Genetic involvement of a cAMP-dependent protein kinase in a G protein signaling pathway regulating morphological and chemical transitions in Aspergillus nidulans. , 2001, Genetics.

[56]  P. Devreotes,et al.  The cAMP receptor family of Dictyostelium. , 1992, International review of cytology.

[57]  L. Cornivelli,et al.  Multiple Regulatory Roles of a Novel Saccharomyces cerevisiae Protein, Encoded by YOL002c, in Lipid and Phosphate Metabolism* , 2002, The Journal of Biological Chemistry.

[58]  D. Natvig,et al.  The nop-1 gene of Neurospora crassa encodes a seven transmembrane helix retinal-binding protein homologous to archaeal rhodopsins. , 1999, Proceedings of the National Academy of Sciences of the United States of America.

[59]  Jae-Hyuk Yu,et al.  The pkaB Gene Encoding the Secondary Protein Kinase A Catalytic Subunit Has a Synthetic Lethal Interaction with pkaA and Plays Overlapping and Opposite Roles in Aspergillus nidulans , 2005, Eukaryotic Cell.