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Role of Trichoderma reesei mitogen-activated protein kinases (MAPKs) in cellulase formation.

ABSTRACT: Despite being the most important cellulase producer, the cellulase-regulating carbon source signal transduction processes in Trichoderma reesei are largely unknown. Elucidating these processes is the key for unveiling how external carbon sources regulate cellulase formation, and ultimately for the improvement of cellulase production and biofuel production from lignocellulose.In this work, the role of the mitogen-activated protein kinase (MAPK) signal transduction pathways on cellulase formation was investigated. The deletion of yeast FUS3-like tmk1 in T. reesei leads to improved growth and significantly improved cellulase formation. However, tmk1 deletion has no effect on the transcription of cellulase-coding genes. The involvement of the cell wall integrity maintenance governing yeast Slt2-like Tmk2 in cellulase formation was investigated by overexpressing tmk3 in T. reesei ?tmk2 to restore cell wall integrity. Transcriptional analysis found little changes in cellulase-coding genes between T. reesei parent, ?tmk2, and ?tmk2::OEtmk3 strains. Cell wall integrity decreased in T. reesei ?tmk2 over the parent strain and restored in ?tmk2::OEtmk3. Meanwhile, cellulase formation is increased in T. reesei ?tmk2 and then decreased in T. reesei ?tmk2::OEtmk3.These investigations elucidate the role of Tmk1 and Tmk2 on cellulase formation: they repress cellulase formation, respectively, by repressing growth and maintaining cell wall integrity, while neither MAPK regulates the transcription of cellulase-coding genes. This work, together with the previous investigations, suggests that all MAPKs are involved in cellulase formation, while Tmk3 is the only MAPK involved in signal transduction for the regulation of cellulase expression on the transcriptional level.

SUBMITTER: Wang M

PROVIDER: S-EPMC5397809 | biostudies-other | 2017

REPOSITORIES: biostudies-other

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Role of <i>Trichoderma reesei</i> mitogen-activated protein kinases (MAPKs) in cellulase formation.

Wang Mingyu M Zhang Meiling M Li Ling L Dong Yanmei Y Jiang Yi Y Liu Kuimei K Zhang Ruiqin R Jiang Baojie B Niu Kangle K Fang Xu X

Biotechnology for biofuels 20170420

<h4>Background</h4>Despite being the most important cellulase producer, the cellulase-regulating carbon source signal transduction processes in <i>Trichoderma reesei</i> are largely unknown. Elucidating these processes is the key for unveiling how external carbon sources regulate cellulase formation, and ultimately for the improvement of cellulase production and biofuel production from lignocellulose.<h4>Results</h4>In this work, the role of the mitogen-activated protein kinase (MAPK) signal tra ...[more]

PMID: 28435444

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Project description:The mitogen-activated protein kinase (MAPK) pathways are important signal transduction pathways conserved in essentially all eukaryotes, but haven't been subjected to functional studies in the most important cellulase-producing filamentous fungus Trichoderma reesei. Previous reports suggested the presence of three MAPKs in T. reesei: Tmk1, Tmk2, and Tmk3. By exploring the phenotypic features of T. reesei ?tmk3, we first showed elevated NaCl sensitivity and repressed transcription of genes involved in glycerol/trehalose biosynthesis under higher osmolarity, suggesting Tmk3 participates in high osmolarity resistance via derepression of genes involved in osmotic stabilizer biosynthesis. We also showed significant downregulation of genes encoding chitin synthases and a ?-1,3-glucan synthase, decreased chitin content, 'budded' hyphal appearance typical to cell wall defective strains, and increased sensitivity to calcofluor white/Congo red in the tmk3 deficient strain, suggesting Tmk3 is involved in cell wall integrity maintenance in T. reesei. We further observed the decrease of cellulase transcription and production in T. reesei ?tmk3 during submerged cultivation, as well as the presence of MAPK phosphorylation sites on known transcription factors involved in cellulase regulation, suggesting Tmk3 is also involved in the regulation of cellulase production. Finally, the expression of cell wall integrity related genes, the expression of cellulase coding genes, cellulase production and biomass accumulation were compared between T. reesei ?tmk3 grown in solid state media and submerged media, showing a strong restoration effect in solid state media from defects resulted from tmk3 deletion. These results showed novel physiological processes that fungal Hog1-type MAPKs are involved in, and present the first experimental investigation of MAPK signaling pathways in T. reesei. Our observations on the restoration effect during solid state cultivation suggest that T. reesei is evolved to favor solid state growth, bringing up the proposal that the submerged condition normally used during investigations on fungal physiology might be misleading.

Dataset Information

Role of Trichoderma reesei mitogen-activated protein kinases (MAPKs) in cellulase formation.

Publications

Role of <i>Trichoderma reesei</i> mitogen-activated protein kinases (MAPKs) in cellulase formation.

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