Cellulase production in filamentous fungi is repressed by various carbon sources. In our preliminary survey in Aspergillus nidulans, degree of de-repression differed depending on carbon sources in a mutant of creA, encoding the transcriptional repressor for carbon catabolite repression (CCR). To further understand mechanisms of CCR of cellulase production, we compared the effects of creA deletion with deletion of protein kinase A (pkaA) and G (ganB) genes, which constitute a nutrient sensing and signaling pathway. In plate culture with carboxymethyl cellulose and d-glucose, deletion of pkaA and ganB, but not creA, led to significant de-repression of cellulase production. In submerged culture with cellobiose and d-glucose or 2-deoxyglucose, both creA or pkaA single deletion led to partial de-repression of cellulase genes with the highest level by their double deletion, while ganB deletion caused de-repression comparable to that of the creA/pkaA double deletion. With ball-milled cellulose and d-glucose, partial de-repression was detected by deletion of creA but not of pkaA or ganB. The creA/pkaA or creA/ganB double deletion led to earlier expression than the creA deletion. Furthermore, the effect of each deletion with d-xylose or L-arabinose as the repressing carbon source was significantly different from that with d-glucose, d-fructose, and d-mannose. Consequently, this study revealed that PkaA and GanB participate in CreA-independent CCR and that contribution of CreA, PkaA, and GanB in CCR differs depending on the inducers, repressing carbon sources, and culture conditions (plate or submerged). Further study of CreA-independent mechanisms is needed to fully understand CCR in filamentous fungi.
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Akada R, Kitagawa T, Kaneko S, Toyonaga D, Ito S, Kakihara Y, Hoshida H, Morimura S, Kondo A, Kida K (2006) PCR-mediated seamless gene deletion and marker recycling in Saccharomyces cerevisiae. Yeast 23:399–405. https://doi.org/10.1002/yea.1365
Alam MA, Kelly JM (2017) Proteins interacting with CreA and CreB in the carbon catabolite repression network in Aspergillus nidulans. Curr Genet 63:669–683. https://doi.org/10.1007/s00294-016-0667-2
Alam MA, Kamlangdee N, Kelly JM (2017) The CreB deubiquitinating enzyme does not directly target the CreA repressor protein in Aspergillus nidulans. Curr Genet 63:647–667. https://doi.org/10.1007/s00294-016-0666-3
Boase NA, Kelly JM (2004) A role for creD, a carbon catabolite repression gene from Aspergillus nidulans, in ubiquitination. Mol Microbiol 53:929–940. https://doi.org/10.1111/j.1365-2958.2004.04172.x
Boase NA, Lockington RA, Adams JR, Rodbourn L, Kelly JM (2003) Molecular characterization and analysis of the acrB gene of Aspergillus nidulans: a gene identified by genetic interaction as a component of the regulatory network that includes the CreB deubiquitination enzyme. Genetics 164:95–104
Brown NA, de Gouvea PF, Krohn NG, Savoldi M, Goldman GH (2013) Functional characterisation of the non-essential protein kinases and phosphatases regulating Aspergillus nidulans hydrolytic enzyme production. Biotechnol Biofuels 6:91. https://doi.org/10.1186/1754-6834-6-91
Brown NA, Dos Reis TF, Ries LN, Caldana C, Mah JH, Yu JH, Macdonald JM, Goldman GH (2015) G-protein coupled receptor-mediated nutrient sensing and developmental control in Aspergillus nidulans. Mol Microbiol 98:420–439. https://doi.org/10.1111/mmi.13135
Chikamatsu G, Shirai K, Kato M, Kobayashi T, Tsukagoshi N (1999) Structure and expression properties of the endo-βAspergillus nidulans. FEMS Microbiol Lett 179:239–245
Colabardini AC, Humanes AC, Gouvea PF, Savoldi M, Goldman MH, Kress MR, Bayram Ö, Oliveira JV, Gomes MD, Braus GH, Goldman GH (2012) Molecular characterization of the Aspergillus nidulans fbxA encoding an F-box protein involved in xylanase induction. Fungal Genet Biol 49:130–140. https://doi.org/10.1016/j.fgb.2011.11.004
Cubero B, Scazzocchio C (1994) Two different, adjacent and divergent zinc finger binding sites are necessary for CREA-mediated carbon catabolite repression in the proline gene cluster of Aspergillus nidulans. EMBO J 13:407–415
Cubero B, Gómez D, Scazzocchio C (2000) Metabolite repression and inducer exclusion in the proline utilization gene cluster of Aspergillus nidulans. J Bacteriol 182:233–235. https://doi.org/10.1128/JB.182.1.233-235.2000
de Souza CP, Hashmi SB, Osmani AH, Andrews P, Ringelberg CS, Dunlap JC, Osmani SA (2013a) Functional analysis of the Aspergillus nidulans kinome. PLoS One 8:e58008. https://doi.org/10.1371/journal.pone.0058008
de Souza WR, Maitan-Alfenas GP, de Gouvêa PF, Brown NA, Savoldi M, Battaglia E, Goldman MH, de Vries RP, Goldman GH (2013b) The influence of Aspergillus niger transcription factors AraR and XlnR in the gene expression during growth in d-xylose, L-arabinose and steam-exploded sugarcane bagasse. Fungal Genet Biol 60:29–45. https://doi.org/10.1016/j.fgb.2013.07.007
de Assis LJ, Ries LN, Savoldi M, Dos Reis TF, Brown NA, Goldman GH (2015) Aspergillus nidulans protein kinase A plays an important role in cellulase production. Biotechnol Biofuels 8:213. https://doi.org/10.1186/s13068-015-0401-1
de Assis LJ, Ulas M, Ries LNA, El Ramli NAM, Sarikaya-Bayram O, Braus GH, Bayram O, Goldman GH (2018) Regulation of Aspergillus nidulans CreA-mediated catabolite repression by the F-Box proteins Fbx23 and Fbx47. MBio 9:e00840–e00818. https://doi.org/10.1128/mBio.00840-18
de la Serna I, Ng D, Tyler BM (1999) Carbon regulation of ribosomal genes in Neurospora crassa occurs by a mechanism which does not require Cre-1, the homologue of the Aspergillus carbon catabolite repressor, CreA. Fungal Genet Biol 26:253–269. https://doi.org/10.1006/fgbi.1999.1121
Dowzer CE, Kelly JM (1989) Cloning of the creA gene from Aspergillus nidulans: a gene involved in carbon catabolite repression. Curr Genet 15:457–459
Dubois M, Gilles K, Hamilton JK, Rebers PA, Smith F (1951) A colorimetric method for the determination of sugars. Nature 168:167. https://doi.org/10.1038/168167a0
Endo Y, Yokoyama M, Morimoto M, Shirai K, Chikamatsu G, Kato N, Tsukagoshi N, Kato M, Kobayashi T (2008) Novel promoter sequence required for inductive expression of the Aspergillus nidulans endoglucanase gene eglA. Biosci Biotechnol Biochem 72:312–320. https://doi.org/10.1271/bbb.70278
Fernändez-Canón JM, Luengo JM (1997) The phenylacetic acid uptake system of Aspergillus nidulans is under a creA-independent model of catabolic repression which seems to be mediated by acetyl-CoA. J Antibiot (Tokyo) 50:45–52. https://doi.org/10.7164/antibiotics.50.45
Gielkens MMC, Dekkers E, Visser J, de Graaff LH (1999) Two cellobiohydrolase-encoding genes from Aspergillus niger require d-xylose and the xylanolytic transcriptional activator XlnR for their expression. Appl Environ Microbiol 65:4340–4345
Hiramoto T, Tanaka M, Ichikawa T, Matsuura Y, Hasegawa-Shiro S, Shintani T, Gomi K (2015) Endocytosis of a maltose permease is induced when amylolytic enzyme production is repressed in Aspergillus oryzae. Fungal Genet Biol 82:136–144. https://doi.org/10.1016/j.fgb.2015.05.015
Ichinose S, Tanaka M, Shintani T, Gomi K (2014) Improved αAspergillus oryzae after a double deletion of genes involved in carbon catabolite repression. Appl Microbiol Biotechnol 98:335–343. https://doi.org/10.1007/s00253-013-5353-4
Ichinose S, Tanaka M, Shintani T, Gomi K (2018) Increased production of biomass-degrading enzymes by double deletion of creA and creB genes involved in carbon catabolite repression in Aspergillus oryzae. J Biosci Bioeng 125:141–147. https://doi.org/10.1016/j.jbiosc.2017.08.019
Ishikawa K, Kunitake E, Kawase T, Atsumi M, Noguchi Y, Ishikawa S, Ogawa M, Koyama Y, Kimura M, Kanamaru K, Kato M, Kobayashi T (2018) Comparison of the paralogous transcription factors AraR and XlnR in Aspergillus oryzae. Curr Genet. https://doi.org/10.1007/s00294-018-0837-5
Kunitake E, Hagiwara D, Miyamoto K, Kanamaru K, Kimura M, Kobayashi T (2016) Regulation of genes encoding cellulolytic enzymes by Pal-PacC signaling in Aspergillus nidulans. Appl Microbiol Biotechnol 100:3621–3635. https://doi.org/10.1007/s00253-016-7409-8
Lafon A, Seo JA, Han KH, Yu JH, d’Enfert C (2005) The heterotrimeric G-protein GanB(alpha)-SfaD(beta)-GpgA(gamma) is a carbon source sensor involved in early cAMP-dependent germination in Aspergillus nidulans. Genetics 171:71–80. https://doi.org/10.1534/genetics.105.040584
Lockington RA, Kelly JM (2002) The WD40-repeat protein CreC interacts with and stabilizes the deubiquitinating enzyme CreB in vivo in Aspergillus nidulans. Mol Microbiol 43:1173–1182
Makita T, Katsuyama Y, Tani S, Suzuki H, Kato N, Todd RB, Hynes MJ, Tsukagoshi N, Kato M, Kobayashi T (2009) Inducer-dependent nuclear localization of a Zn(II)2Cys6 transcriptional activator, AmyR, in Aspergillus nidulans. Biosci Biotechnol Biochem 73:391–399. https://doi.org/10.1271/bbb.80654
Marui J, Kitamoto N, Kato M, Kobayashi T, Tsukagoshi N (2002) Transcriptional activator, AoXlnR, mediates cellulose-inductive expression of the xylanolytic and cellulolytic genes in Aspergillus oryzae. FEBS Lett 528:279–282
Noguchi Y, Sano M, Kanamaru K, Ko T, Takeuchi M, Kato M, Kobayashi T (2009) Genes regulated by AoXlnR, the xylanolytic and cellulolytic transcriptional regulator, in Aspergillus oryzae. Appl Microbiol Biotechnol 85:141–154. https://doi.org/10.1007/s00253-009-2236-9
Noguchi Y, Tanaka H, Kanamaru K, Kato M, Kobayashi T (2011) Xylose triggers reversible phosphorylation of XlnR, the fungal transcriptional activator of xylanolytic and cellulolytic genes in Aspergillus oryzae. Biosci Biotechnol Biochem 75:953–959. https://doi.org/10.1271/bbb.100923
Orejas M, MacCabe AP, Pérez-González JA, Kumar S, Ramón D (2001) The wide-domain carbon catabolite repressor CreA indirectly controls expression of the Aspergillus nidulans xlnB gene, encoding the acidic endo-β
Ries LN, Beattie SR, Espeso EA, Cramer RA, Goldman GH (2016) Diverse regulation of the CreA carbon catabolite repressor in Aspergillus nidulans. Genetics 203:335–352. https://doi.org/10.1534/genetics.116.187872
Rowlands RT, Turner G (1973) Nuclear and extranuclear inheritance of oligomycin resistance in Aspergillus nidulans. Mol Gen Genet 126:201–216
Shroff RA, Lockington RA, Kelly JM (1996) Analysis of mutations in the creA gene involved in carbon catabolite repression in Aspergillus nidulans. Can J Microbiol 42:950–959
Somogyi M (1952) Notes on sugar determination. J Biol Chem 195:19–23
Strauss J, Mach RL, Zeilinger S, Hartler G, Stöffler G, Wolschek M, Kubicek CP (1995) Cre1, the carbon catabolite repressor protein from Trichoderma reesei. FEBS Lett 376:103–107
Tamayo EN, Villanueva A, Hasper AA, de Graaff LH, Ramón D, Orejas M (2008) CreA mediates repression of the regulatory gene xlnR which controls the production of xylanolytic enzymes in Aspergillus nidulans. Fungal Genet Biol 45:984–993. https://doi.org/10.1016/j.fgb.2008.03.002
Tamayo-Ramos JA, Flipphi M, Pardo E, Manzanares P, Orejas M (2012) L-rhamnose induction of Aspergillus nidulansα
Tanaka M, Ichinose S, Shintani T, Gomi K (2018) Nuclear export-dependent degradation of the carbon catabolite repressor CreA is regulated by a region located near the C-terminus in Aspergillus oryzae. Mol Microbiol. https://doi.org/10.1111/mmi.14072
van Peij NN, Gielkens MM, de Vries RP, Visser J, de Graaff LH (1998) The transcriptional activator XlnR regulates both xylanolytic and endoglucanase gene expression in Aspergillus niger. Appl Environ Microbiol 64:3615–3619
Vega M, Riera A, Fernández-Cid A, Herrero P, Moreno F (2016) Hexokinase 2 is an intracellular glucose sensor of yeast cells that maintains the structure and activity of Mig1 protein repressor complex. J Biol Chem 291:7267–7285. https://doi.org/10.1074/jbc.M115.711408
This work was partially supported by the Program for Promotion of Basic and Applied Research for Innovations in Bio-oriented Industry and by the Science and Technology Research Promotion Program for Agriculture, Forestry, Fisheries, and Food Industry. It was also supported by JSPS KAKENHI (Grant Numbers JP 18H02125 and JP 17H06763) and funding from Institute for Fermentation, Osaka (IFO) and Noda Institute for Scientific Research.
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Kunitake, E., Li, Y., Uchida, R. et al. CreA-independent carbon catabolite repression of cellulase genes by trimeric G-protein and protein kinase A in Aspergillus nidulans. Curr Genet 65, 941–952 (2019). https://doi.org/10.1007/s00294-019-00944-4
- Aspergillus nidulans
- Carbon catabolite repression