Abstract
Trichoderma reesei is the most important industrial cellulase-producing filamentous fungus. Although its molecular physiology has been investigated, the signal transduction pathways are not fully understood. In particular, the role of casein kinase II (CKII) is not yet clear. In this work, we carried out functional investigations on a catalytic subunit of CKII, CKIIα2. Comparison of the phenotypic features of T. reesei parent and Δck2α2 strains showed significant changes following ck2α2 disruption. T. reesei Δck2α2 form significantly smaller mycelial pellets in glucose-containing liquid minimum media, have shorter and fewer branch hyphae, produce smaller amounts of chitinases, produce more spores, show more robust growth on glucose-containing agar plates, and consume glucose at a significantly higher rate. Suggestions can be made that CKIIα2 governs chitinase expression, and the disruption of ck2α2 results in lower levels of chitinase production, leading to a weaker cell wall disruption capability, further resulting in weaker hyphal branching, which eventually leads to smaller mycelial pellets in liquid media. Further conclusions can be made that CKIIα2 is involved in repression of sporulation and glucose metabolism, which is consistent with the proposal that CKIIα2 represses global metabolism. These observations make the deletion of ck2α2 a potentially beneficial genetic disruption for T. reesei during industrial applications, as smaller mycelial pellets, more spores and more robust glucose metabolism are all desired traits for industrial fermentation. This work reports novel unique functions of a CKII catalytic subunit and is also the first genetic and physiological investigation on CKII in T. reesei.
Similar content being viewed by others
References
Abramczyk O, Zień P, Zieliński F, Pilecki M, Hellman U, Szyszka R (2003) The protein kinase 60S is a free catalytic CK2α' subunit and forms an inactive complex with superoxide dismutase SOD1. Biochem Biophys Res Commun 307:31–40. doi:10.1016/S0006-291X(03)01126-4
Bidwai AP, Reed JC, Glover CV (1995) Cloning and disruption of CKB1, the gene encoding the 38-kDa β subunit of Saccharomyces cerevisiae casein kinase II (CKII): deletion of CKII regulatory subunits elicits a salt-sensitive phenotype. J Biol Chem 270(18):10395–10404. doi:10.1074/jbc.270.18.10395
Cherry JR, Fidantsef AL (2003) Directed evolution of industrial enzymes: an update. Curr Opin Biotechnol 14(4):438–443. doi:10.1016/S0958-1669(03)00099-5
Cziferszky A, Mach RL, Kubicek CP (2002) Phosphorylation positively regulates DNA binding of the carbon catabolite repressor Cre1 of Hypocrea jecorina (Trichoderma reesei). J Biol Chem 277(17):14688–14694. doi:10.1074/jbc.M200744200
Domańska K, Zieliński R, Kubiński K, Sajnaga E, Maslyk M, Bretner M, Szyszka R (2005) Different properties of four molecular forms of protein kinase CK2 from Saccharomyces cerevisiae. Acta Biochim Pol 52(4):947–951
Gruber F, Visser J, Kubicek CP, de Graaff LH (1990) The development of a heterologous transformation system for the cellulolytic fungus Trichoderma reesei based on a pyrG-negative mutant strain. Curr Genet 18(1):71–76. doi:10.1007/BF00321118
He Q, Cha J, He Q, Lee HC, Yang Y, Liu Y (2006) CKI and CKII mediate the FREQUENCY-dependent phosphorylation of the WHITE COLLAR complex to close the Neurospora circadian negative feedback loop. Genes Dev 20(18):2552–2565. doi:10.1101/gad.1463506
Hideno A, Inoue H, Tsukahara K, Yano S, Fang X, Endo T, Sawayama S (2011) Production and characterization of cellulases and hemicellulases by Acremonium cellulolyticus using rice straw subjected to various pretreatments as the carbon source. Enzym Microb Technol 48(2):162–168. doi:10.1016/j.enzmictec.2010.10.005
Ilmén M, Saloheimo A, Onnela ML, Penttilä ME (1997) Regulation of cellulase gene expression in the filamentous fungus Trichoderma reesei. Appl Environ Microbiol 63(4):1298–1306
Ivanova C, Baath JA, Seiboth B, Kubicek CP (2013) Systems analysis of lactose metabolism in Trichoderma reesei identifies a lactose permease that is essential for cellulase induction. PLoS ONE 8(5):e62631. doi:10.1371/journal.pone.0062631
Kubicek CP, Mikus M, Schuster A, Schmoll M, Seiboth B (2009) Metabolic engineering strategies for the improvement of cellulase production by Hypocrea jecorina. Biotechnol Biofuels 2:19. doi:10.1186/1754-6834-2-19
Larkin MA, Blackshields G, Brown NP, Chenna R, McGettigan PA, McWilliam H, Valentin F, Wallace IM, Wilm A, Lopez R, Thompson JD, Gibson TJ, Higgins DG (2007) Clustal W and Clustal X version 2.0. Bioinformatics 23(21):2947–2948. doi:10.1093/bioinformatics/btm404
Lee Y, Park I, Yoo J, Chung S, Lee Y, Cho Y, Ahn S, Kim C, Choi Y (2007) Cloning, purification, and characterization of chitinase from Bacillus sp. DAU101. Bioresour Technol 98:2734–2741. doi:10.1016/j.biortech.2006.09.048
Lei Y, Liu G, Li Z, Gao L, Qin Y, Qu Y (2014) Functional characterization of protein kinase CK2 regulatory subunits regulating Penicillium oxalicum asexual development and hydrolytic enzyme production. Fungal Genet Biol 66:44–53. doi:10.1016/j.fgb.2014.02.007
Lin Y, Tanaka S (2006) Ethanol fermentation from biomass resources: current state and prospects. Appl Microbiol Biotechnol 69(6):627–642. doi:10.1007/s00253-005-0229-x
Litchfield DW (2003) Protein kinase CK2: structure, regulation and role in cellular decisions of life and death. Biochem J 369(Pt 1):1–15. doi:10.1042/BJ20021469
Lowry OH, Rosebrough NJ, Farr AL, Randall RJ (1951) Protein measurement with the Folin phenol reagent. J Biol Chem 193(1):265–275
Martinez D, Berka RM, Henrissat B, Saloheimo M, Arvas M, Baker SE, Chapman J, Chertkov O, Coutinho PM, Cullen D, Danchin EG, Grigoriev IV, Harris P, Jackson M, Kubicek CP, Han CS, Ho I, Larrondo LF, de Leon AL, Magnuson JK, Merino S, Misra M, Nelson B, Putnam N, Robbertse B, Salamov AA, Schmoll M, Terry A, Thayer N, Westerholm-Parvinen A, Schoch CL, Yao J, Barabote R, Nelson MA, Detter C, Bruce D, Kuske CR, Xie G, Richardson P, Rokhsar DS, Lucas SM, Rubin EM, Dunn-Coleman N, Ward M, Brettin TS (2008) Genome sequencing and analysis of the biomass-degrading fungus Trichoderma reesei (syn. Hypocrea jecorina). Nat Biotechnol 26(5):553–560. doi:10.1038/nbt1403
Meggio F, Pinna LA (2003) One-thousand-and-one substrates of protein kinase CK2? FASEB J 17(3):349–368. doi:10.1096/fj.02-0473rev
Mehra A, Shi M, Baker CL, Colot HV, Loros JJ, Dunlap JC (2009) A role for casein kinase 2 in the mechanism underlying circadian temperature compensation. Cell 137(4):749–760. doi:10.1016/j.cell.2009.03.019
Montenecourt BS, Eveleigh DE (1977) Preparation of mutants of Trichoderma reesei with enhanced cellulase production. Appl Environ Microbiol 34(6):777–782
Papagianni M (2004) Fungal morphology and metabolite production in submerged mycelial processes. Biotechnol Adv 22(3):189–259. doi:10.1016/j.biotechadv.2003.09.005
Salamon JA, Acuna R, Dawe AL (2010) Phosphorylation of phosducin-like protein BDM-1 by protein kinase 2 (CK2) is required for virulence and Gβ subunit stability in the fungal plant pathogen Cryphonectria parasitica. Mol Microbiol 76(4):848–860. doi:10.1111/j.1365-2958.2010.07053.x
Schmoll M (2008) The information highways of a biotechnological workhorse-signal transduction in Hypocrea jecorina. BMC Genomics 9:430. doi:10.1186/1471-2164-9-430
Schneider CA, Rasband WS, Eliceiri KW (2012) NIH Image to ImageJ: 25 years of image analysis. Nat Methods 9(7):671–675. doi:10.1038/nmeth.2089
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(7):2168–2178. doi:10.1128/AEM. 06959-11
Seibel C, Tisch D, Kubicek CP, Schmoll M (2012) ENVOY is a major determinant in regulation of sexual development in Hypocrea jecorina (Trichoderma reesei). Eukaryot Cell 11(7):885–895. doi:10.1128/EC.05321-11
Sternberg D, Mandels GR (1979) Induction of cellulolytic enzymes in Trichoderma reesei by sophorose. J Bacteriol 139(3):761–769
Tisch D, Schmoll M (2013) Targets of light signalling in Trichoderma reesei. BMC Genomics 14:657. doi:10.1186/1471-2164-14-657
Tripodi F, Zinzalla V, Vanoni M, Alberghina L, Coccetti P (2007) In CK2 inactivated cells the cyclin dependent kinase inhibitor Sic1 is involved in cell-cycle arrest before the onset of S phase. Biochem Biophys Res Commun 359(4):921–927. doi:10.1016/j.bbrc.2007.05.195
Wang M, Li Z, Fang X, Wang L, Qu Y (2012) Cellulolytic enzyme production and enzymatic hydrolysis for second-generation bioethanol production. Adv Biochem Eng Biotechnol 128:1–24. doi:10.1007/10_2011_131
Wang F, Liang Y, Wang M, Yang H, Liu K, Zhao Q, Fang X (2013a) Functional diversity of the p24γ homologue Erp reveals physiological differences between two filamentous fungi. Fungal Genet Biol 61:15–22. doi:10.1016/j.fgb.2013.08.017
Wang M, Zhao Q, Yang J, Jiang B, Wang F, Liu K, Fang X (2013b) A mitogen-activated protein kinase Tmk3 participates in high osmolarity resistance, cell wall integrity maintenance and cellulase production regulation in Trichoderma reesei. PLoS ONE 8(8):e72189. doi:10.1371/journal.pone.0072189
Wang M, Dong Y, Zhao Q, Wang F, Liu K, Jiang B, Fang X (2014) Identification of the role of a MAP kinase Tmk2 in Hypocrea jecorina (Trichoderma reesei). Sci Rep 4:6732. doi:10.1038/srep06732
Yamazaki H, Tanaka A, Kaneko J, Ohta A, Horiuchi H (2008) Aspergillus nidulans ChiA is a glycosylphosphatidylinositol (GPI)-anchored chitinase specifically localized at polarized growth sites. Fungal Genet Biol 45(6):963–972. doi:10.1016/j.fgb.2008.02.008
Yang Y, Cheng P, Liu Y (2002) Regulation of the Neurospora circadian clock by casein kinase II. Genes Dev 16(8):994–1006. doi:10.1101/gad.965102
Zhang G, Hartl L, Schuster A, Polak S, Schmoll M, Wang T, Seidl V, Seiboth B (2009) Gene targeting in a nonhomologous end joining deficient Hypocrea jecorina. J Biotechnol 139(2):146–151. doi:10.1016/j.jbiotec.2008.10.007
Zhang J, Zhang Y, Zhong Y, Qu Y, Wang T (2012) Ras GTPases modulate morphogenesis, sporulation and cellulase gene expression in the cellulolytic fungus Trichoderma reesei. PLoS ONE 7(11):e48786. doi:10.1371/journal.pone.0048786
Acknowledgments
This work was supported by the National Key Technology R&D Program of China (No. 2014BAD02B07), National Energy Applied Technology Research and Demonstration Project (No. NY20130402), National Natural Science Foundation of China (No. 31200051), Shandong Province Natural Science Foundation (No. ZR2012CQ022 and No. ZR2013CM041) and Scientific Research Foundation for the Returned Overseas Chinese Scholars, State Education Ministry.
Author information
Authors and Affiliations
Corresponding author
Electronic supplementary material
Below is the link to the electronic supplementary material.
ESM 1
(PDF 675 kb)
Rights and permissions
About this article
Cite this article
Wang, M., Yang, H., Zhang, M. et al. Functional analysis of Trichoderma reesei CKIIα2, a catalytic subunit of casein kinase II. Appl Microbiol Biotechnol 99, 5929–5938 (2015). https://doi.org/10.1007/s00253-015-6544-y
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00253-015-6544-y