Abstract
Trichoderma serves as the primary producer of cellulases and hemicellulases in industrial settings as it readily secretes a variety of cellulolytic enzymes. The protein kinase SNF1 (sucrose-nonfermenting 1) can enable cells to adapt to changes in carbon metabolism by phosphorylating key rate-limiting enzymes involved in the maintenance of energy homeostasis and carbon metabolism within cells. Histone acetylation is an important epigenetic regulatory mechanism that influences physiological and biochemical processes. GCN5 is a representative histone acetylase involved in promoter chromatin remodeling and associated transcriptional activation. Here, the TvSNF1 and TvGCN5 genes were identified in Trichoderma viride Tv-1511, which exhibits promising activity with respect to its ability to produce cellulolytic enzymes for biological transformation. The SNF1-mediated activation of the histone acetyltransferase GCN5 was herein found to promote cellulase production in T. viride Tv-1511 via facilitating changes in histone acetylation. These results demonstrated that cellulolytic enzyme activity and the expression of genes encoding cellulases and transcriptional activators were clearly enhanced in T. viride Tv-1511 mutants in which TvSNF1 and TvGCN5 were overexpressed, with concomitant changes in histone H3 acetylation levels associated with these genes. GCN5 was also found to be directly recruited to promoter regions to alter histone acetylation, while SNF1 functioned upstream as a transcriptional activator that promotes GCN5 upregulation at the mRNA and protein levels in the context of cellulase induction in T. viride Tv-1511. These findings underscore the important role that this SNF1-GCN5 cascade plays in regulating cellulase production in T. viride Tv-1511 by promoting altered histone acetylation, offering a theoretical basis for the optimization of T. viride in the context of industrial cellulolytic enzyme production.
Key points
• SNF1 kinase and GCN5 acetylase promoted cellulase production in Trichoderma by increasing the expression of genes encoding cellulases and transcriptional activators
• SNF1 and GCN5 promoted cellulase production by driving H3ac modifications, and GCN5 directly band to the promoter regions to catalyze distinct H3ac modifications
• SNF1 acts upstream of GCN5 as a transcriptional activator in the cellulase production of Trichoderma
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All data generated or analyzed during this study are included in this published article (and its supplementary information files).
References
Abate G, Bastonini E, Braun K, Verdone L, Young E, Caserta M (2012) Snf1/AMPK regulates Gcn5 occupancy, H3 acetylation and chromatin remodeling at S. cerevisiae ADY2 promoter. BBA-Gene Regul Mech 1819:419–427. https://doi.org/10.1016/j.bbagrm.2012.01.009
Aghcheh RK, Kubicek CP (2015) Epigenetics as an emerging tool for improvement of fungal strains used in biotechnology. Appl Microbiol Biotechnol 99:6167–6181. https://doi.org/10.1007/s00253-015-6763-2
Antoniêto ACC, de Paula RG, Castro LDS, Silva-Rocha R, Persinoti GF, Silva RN (2016) Trichoderma reesei CRE1-mediated carbon catabolite repression in response to sophorose through RNA sequencing analysis. Curr Genom 17:119–131. https://doi.org/10.2174/1389202917666151116212901
Benocci T, Aguilar-Pontes MV, Zhou MM, Seiboth B, de Vries RP (2017) Regulators of plant biomass degradation in ascomycetous fungi. Biotechnol Biofuels 10:152. https://doi.org/10.1186/s13068-017-0841-x
Bischof RH, Ramoni J, Seiboth B (2016) Cellulases and beyond: the first 70 years of the enzyme producer Trichoderma reesei. Microb Cell Fact 15:106. https://doi.org/10.1186/s12934-016-0507-6
Broach JR (2012) Nutritional control of growth and development in yeast. Genetics 192:73–105. https://doi.org/10.1534/genetics.111.135731
Cánovas D, Marcos AT, Gacek A, Ramos MS, Strauss J (2014) The histone acetyltransferase GcnE (GCN5) plays a central role in the regulation of Aspergillus asexual development. Genetics 197:1175–1189. https://doi.org/10.1534/genetics.114.165688
Castro LDS, Antoniêto ACC, Pedersoli WR, Silva-Rocha R, Persinoti GF, Silva RN (2014) Expression pattern of cellulolytic and xylanolytic genes regulated by transcriptional factors XYR1 and CRE1 are affected by carbon source in Trichoderma reesei. Gene Expr Patterns 14:88–95. https://doi.org/10.1016/j.gep.2014.01.003
Charbon G, Breunig KD, Wattiez R, Vandenhaute J, Noel-Georis I (2004) Key role of Ser562/661 in Snf1-dependent regulation of Cat8p in Saccharomyces cerevisiae and Kluyveromyces lactis. Mol Cell Biol 24:4083–4091. https://doi.org/10.1128/MCB.24.10.4083-4091.2004
Cheng Y, Song X, Qin Y, Qu Y (2009) Genome shuffling improves production of cellulase by Penicillium decumbens JU-A10. J Appl Microbiol 107:1837–1846. https://doi.org/10.1111/j.1365-2672.2009.04362.x
de Assis LJ, Silva LP, Bayram O, Dowling P, Kniemeyer O, Krüger T, Brakhage AA, Chen Y, Dong L, Tan K (2021) Carbon catabolite repression in filamentous fungi is regulated by phosphorylation of the transcription factor CreA. mBio 12:e03146. https://doi.org/10.1128/mBio.03146-20
de Paula RG, Antoniêto ACC, Ribeiro LFC, Srivastava N, O’Donovan A, Mishra PK, Gupta VK, Silva RN (2019) Engineered microbial host selection for value-added bioproducts from lignocellulose. Biotechnol Adv 37:107347. https://doi.org/10.1016/j.biotechadv.2019.02.003
de Risi JL, Iyer VR, Brown PO (1997) Exploring the metabolic and genetic control of gene expression on a genomic scale. Science 278:680–686. https://doi.org/10.1126/science.278.5338.680
Fan A, Mi W, Liu Z, Zeng G, Zhang P, Hu Y (2017) Deletion of a histone acetyltransferase leads to the pleiotropic activation of natural products in Metarhizium robertsii. Org Lett 19:1686–1689. https://doi.org/10.1021/acs.orglett.7b00476
Friedl MA, Schmoll M, Kubicek CP, Druzhinina IS (2008) Photostimulation of Hypocrea atroviridis growth occurs due to a cross-talk of carbon metabolism, blue light receptors and response to oxidative stress. Microbiology 154:1229–1241. https://doi.org/10.1099/mic.0.2007/014175-0
Hansen W, Yourassowsky E (1984) Detection of beta-glucuronidase in lactose-fermenting members of the family Enterobacteriaceae and its presence in bacterial urine cultures. J Clin Microbiol 20:1177–1179. https://doi.org/10.1128/jcm.20.6.1177-1179.1984
Hardie DG (2007) AMP-activated/SNF1 protein kinases: conserved guardians of cellular energy. Nat Rev Mol Cell Biol 8:774–785. https://doi.org/10.1038/nrm2249
Hedbacker K, Carlson M (2008) SNF1/AMPK pathways in yeast. Front Biosci 13:2408–2420. https://doi.org/10.2741/2854
Hu YR, Hu SS, Xu WZ, Zhu J, Shi L, Ren A, Zhao MW (2020) In Ganoderma lucidum, Glsnf1 regulates cellulose degradation by inhibiting GlCreA during the utilization of cellulose. Environ Microbiol 22:107–121. https://doi.org/10.1111/1462-2920.14826
Jenuwein T, Allis CD (2001) Translating the histone code. Science 293:1074–1080. https://doi.org/10.1126/science.1063127
Kishkevich A, Cooke SL, Harris MRA, Bruin RD (2019) Gcn5 and Rpd3 have a limited role in the regulation of cell cycle transcripts during the G1 and S phases in Saccharomyces cerevisiae. Sci Rep 9:10686. https://doi.org/10.1038/s41598-019-47170-z
Lee KK, Workman JL (2007) Histone acetyltransferase complexes: one size doesn’t fit all. Nat Rev Mol Cell Biol 8:284–295. https://doi.org/10.1038/nrm2145
Lesage P, Yang X, Carlson M (1996) Yeast Snf1 protein kinase interacts with Sip4, a C6 zinc cluster transcriptional activator: a new role for Snf1 in the glucose response. Mol Cell Biol 16: 1921–1928. 0270–7306/96/$04.00+0
Li Z, Zhang H, Cai CJ, Lin Z, Zhen Z, Chu J, Guo K (2022) Histone acetyltransferase GCN5-mediated lysine acetylation modulates salt stress adaption of Trichoderma. Appl Microbiol Bio 106:3033–3049. https://doi.org/10.1007/s00253-022-11897-z
Liu Y, Xu X, Kuo MH (2010) Snf1p regulates Gcn5p transcriptional activity by antagonizing Spt3p. Genetics 184:91–105. https://doi.org/10.1534/genetics.109.110957
Liu KM, Dong YM, Wang FZ, Jiang BJ, Wang MY, Fang X (2016) Regulation of cellulase expression, sporulation, and morphogenesis by velvet family proteins in Trichoderma reesei. Appl Microbiol Biotechnol 100:769–779. https://doi.org/10.1007/s00253-015-7059-2
Marcel N, Boysen JM, Thomas K, Knig CC, Falk H, Thomas M, Brakhage AA (2021) Yeast two-hybrid screening reveals a dual function for the histone acetyltransferase GcnE by controlling glutamine synthesis and development in Aspergillus fumigatus. Curr Genet 65:523–538. https://doi.org/10.1007/s00294-018-0891-z
Marmorstein R, Zhou MM (2014) Writers and readers of histone acetylation: structure, mechanism, and inhibition. CSH Persp Biol 6:a018762. https://doi.org/10.1101/cshperspect.a018762
Mattam AJ, Chaudhari YB, Velankar HR (2022) Factors regulating cellulolytic gene expression in filamentous fungi: an overview. Microb Cell Fact 21:1–19. https://doi.org/10.1186/s12934-022-01953-8
Mutlu B, Puigserver P (2020) Gcn5 acetyltransferase in cellular energetic and metabolic processes. BBA Gene Regul Mech 1864:194626. https://doi.org/10.1016/j.bbagrm.2020.194626
Nadal M, Garcia-Pedrajas MD, Gold SE (2010) The snf1 gene of Ustilago maydis acts as a dual regulator of cell wall degrading enzymes. Phytopathology 100:1364–1372. https://doi.org/10.1094/PHYTO-01-10-0011
Ospina-Giraldo MD, Mullins E, Kang S (2003) Loss of function of the Fusarium oxysporum SNF1 gene reduces virulence on cabbage and Arabidopsis. Curr Genet 44:49–57. https://doi.org/10.1007/s00294-003-0419-y
Ostling J, Ronne H (1998) Negative control of the Mig1p repressor by Snf1-dependent phosphorylation in the absence of glucose. Eur J Biochem 252:162–168. https://doi.org/10.1046/j.1432-1327.1998.2520162.x
Poças-Fonseca MJ, Cabral CG, Manfrão-Netto JHC (2020) Epigenetic manipulation of filamentous fungi for biotechnological applications: a systematic review. Biotechnol Lett 42:885–904. https://doi.org/10.1007/s10529-020-02871-8
Raab AM, Hlavacek V, Bolotina N, Lang C (2011) Shifting the fermentative/oxidative balance in Saccharomyces cerevisiae by transcriptional deregulation of Snf1 via overexpression of the upstream activating kinase Sak1p. Appl Environ Microbiol 77:1981–1989. https://doi.org/10.1128/AEM.02219-10
Ramírez-Zavala B, Mottola A, Haubenreißer J, Schneider S, Allert S, Brunke S, Ohlsen K, Hube B, Morschhäuser J (2017) The Snf1-activating kinase Sak1 is a key regulator of metabolic adaptation and in vivo fitness of Candida albicans. Mol Microbiol 104:989–1007. https://doi.org/10.1111/mmi.13674
Schuster A, Tisch D, Seidl-Seiboth V, Kubicek CP, Schmoll M (2012) Roles of protein kinase A and adenylate cyclase in light-modulated cellulase regulation in Trichoderma reesei. Appl Environ Microbiol 78:2168–2178. https://doi.org/10.1128/Aem.06959-11
Shirra MK, Rogers SE, Alexander DE, Arndt K (2005) The Snf1 protein kinase and Sit4 protein phosphatase have opposing functions in regulating TATA-binding protein association with the Saccharomyces cerevisiae INO1 promoter. Genetics 169:1957–1972. https://doi.org/10.1534/genetics.104.038075
Stefano B, Patrizia B, Matteo C, Massimo G (2016) Inverse PCR and quantitative PCR as alternative methods to southern blotting analysis to assess transgene copy number and characterize the integration site in transgenic woody plants. Biochem Genet 54:291–305
Tonukari NJ, Scott-Craig JS, Walton JD (2000) The Cochliobolus carbonum SNF1 gene is required for cell wall-degrading enzyme expression and virulence on maize. Plant Cell 12:237–247. https://doi.org/10.1105/tpc.12.2.237
Wang L, Mizzen C, Ying C, Candau R, Barlev N, Brownell J, Allis CD, Berger SL (1997) Histone acetyltransferase activity is conserved between yeast and human GCN5 and is required for complementation of growth and transcriptional activation. Mol Cell Biol 17:519–527. https://doi.org/10.1128/MCB.17.1.519
Xin Q, Gong Y, Lv X, Chen G, Liu W (2013) Trichoderma reesei histone acetyltransferase GCN5 regulates fungal growth, conidiation, and cellulase gene expression. Curr Microbiol 67:580–589. https://doi.org/10.1007/s00284-013-0396-4
Young ET, Kacherovsky N, van Riper K (2002) Snf1 protein kinase regulates Adr1 binding to chromatin but not transcription activation. J Biol Chem 277:38095–38103. https://doi.org/10.1074/jbc.M206158200
Zhang JJ, Zhang GX, Wang W, Wang W, Wei DZ (2018) Enhanced cellulase production in Trichoderma reesei RUT C30 via constitution of minimal transcriptional activators. Microb Cell Fact 17:75. https://doi.org/10.1186/s12934-018-0926-7
Zhao S, Zhang X, Li HT (2018) Beyond histone acetylation-writing and erasing histone acetylations. Curr Opin Struc Biol 53:169–177. https://doi.org/10.1016/j.sbi.2018.10.001
Funding
This research is supported by the Natural Science Foundation of Shandong Province (ZR202102270296), the Education and Industry Integration Innovation Pilot Project of Qilu University of Technology (Shandong Academy of Sciences) (No. 2022JBZ01-06), the Program for Innovation Team in the University of Jinan (2019GXRC033), the National Natural Science Foundation of China (21607169; 41977125; 41907033), the Natural Science Foundation of Guangdong Province (2019A1515011948), the Foundation of International Technology Cooperation Project from Qilu University of Technology (Shandong Academy of Sciences) (2022GH026), and the Youth Science Funds of Shandong Academy of Sciences (2020QN0019).
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ZL conceived the project; ZL and ZHL designed the experiments; ZL, CJC, XXH and XL carried out the experiments; ZL and CJC analyzed the data; ZL and ZHL wrote the article. All authors read and approved the final manuscript.
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Li, Z., Cai, C., Huo, X. et al. Sucrose-nonfermenting 1 kinase activates histone acetylase GCN5 to promote cellulase production in Trichoderma. Appl Microbiol Biotechnol 107, 4917–4930 (2023). https://doi.org/10.1007/s00253-023-12617-x
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DOI: https://doi.org/10.1007/s00253-023-12617-x