STK-12 acts as a transcriptional brake to control the expression of cellulase-encoding genes in Neurospora crassa


Autoři: Liangcai Lin aff001;  Shanshan Wang aff001;  Xiaolin Li aff001;  Qun He aff002;  J. Philipp Benz aff003;  Chaoguang Tian aff001
Působiště autorů: Key Laboratory of Systems Microbial Biotechnology, Tianjin Institute of Industrial Biotechnology, Chinese Academy of Sciences, Tianjin, China aff001;  State Key Laboratory of Agrobiotechnology and MOA Key Laboratory of Soil Microbiology, College of Biological Sciences, China Agricultural University, Beijing, China aff002;  Technical University of Munich, TUM School of Life Sciences Weihenstephan, Hans-Carl-von-Carlowitz-Platz, Freising, Germany aff003;  Technical University of Munich, Institute for Advanced Study, Lichtenbergstr, Garching, Germany aff004
Vyšlo v časopise: STK-12 acts as a transcriptional brake to control the expression of cellulase-encoding genes in Neurospora crassa. PLoS Genet 15(11): e32767. doi:10.1371/journal.pgen.1008510
Kategorie: Research Article
doi: 10.1371/journal.pgen.1008510

Souhrn

Cellulolytic fungi have evolved a complex regulatory network to maintain the precise balance of nutrients required for growth and hydrolytic enzyme production. When fungi are exposed to cellulose, the transcript levels of cellulase genes rapidly increase and then decline. However, the mechanisms underlying this bell-shaped expression pattern are unclear. We systematically screened a protein kinase deletion set in the filamentous fungus Neurospora crassa to search for mutants exhibiting aberrant expression patterns of cellulase genes. We observed that the loss of stk-12 (NCU07378) caused a dramatic increase in cellulase production and an extended period of high transcript abundance of major cellulase genes. These results suggested that stk-12 plays a critical role as a brake to turn down the transcription of cellulase genes to repress the overexpression of hydrolytic enzymes and prevent energy wastage. Transcriptional profiling analyses revealed that cellulase gene expression levels were maintained at high levels for 56 h in the Δstk-12 mutant, compared to only 8 h in the wild-type (WT) strain. After growth on cellulose for 3 days, the transcript levels of cellulase genes in the Δstk-12 mutant were 3.3-fold over WT, and clr-2 (encoding a transcriptional activator) was up-regulated in Δstk-12 while res-1 and rca-1 (encoding two cellulase repressors) were down-regulated. Consequently, total cellulase production in the Δstk-12 mutant was 7-fold higher than in the WT. These results strongly suggest that stk-12 deletion results in dysregulation of the cellulase expression machinery. Further analyses showed that STK-12 directly targets IGO-1 to regulate cellulase production. The TORC1 pathway promoted cellulase production, at least partly, by inhibiting STK-12 function, and STK-12 and CRE-1 functioned in parallel pathways to repress cellulase gene expression. Our results clarify how cellulase genes are repressed at the transcriptional level during cellulose induction, and highlight a new strategy to improve industrial fungal strains.

Klíčová slova:

Cellulases – Fungal genetics – Fungi – Gene expression – Gene regulation – Neurospora crassa – Phosphorylation – Saccharomyces cerevisiae


Zdroje

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