Abstract

BackgroundThe canonical heterotrimeric G protein-cAMP/PKA pathway regulates numerous cellular processes in filamentous fungi. Chaetomium globosum, a saprophytic fungus, is known for producing many secondary metabolites, including cytotoxic chaetoglobosin A (ChA), as well as abundant cellulase and xylanase.ResultsHere we report on the functional characterization of this signaling pathway in C. globosum. We blocked the pathway by knocking down the putative Gα-encoding gene gna1 (in the pG14 mutant). This led to impaired cellulase production and significantly decreased transcription of the major cellulase and xylanase genes. Almost all the glycohydrolase family genes involved in cellulose degradation were downregulated, including the major cellulase genes, cel7a, cel6a, egl1, and egl2. Importantly, the expression of transcription factors was also found to be regulated by gna1, especially Ace1, Clr1/2 and Hap2/3/5 complex. Additionally, carbon metabolic processes including the starch and sucrose metabolism pathway were substantially diminished, as evidenced by RNA-Seq profiling and quantitative reverse transcription (qRT)-PCR. Interestingly, these defects could be restored by simultaneous knockdown of the pkaR gene encoding the regulatory subunit of cAMP-dependent PKA (in the pGP6 mutant) or supplement of the cAMP analog, 8-Br-cAMP. Moreover, the Gα-cAMP/PKA pathway regulating cellulase production is modulated by environmental signals including carbon sources and light, in which VelB/VeA/LaeA complex and ENVOY probably work as downstream effectors.ConclusionThese results revealed, for the first time, the positive role of the heterotrimeric Gα-cAMP/PKA pathway in the regulation of cellulase and xylanase utilization in C. globosum.

Highlights

  • The canonical heterotrimeric G protein-cyclic adenosine monophosphate (cAMP)/cAMP-dependent protein kinase catalytic subunit A (PKA) pathway regulates numerous cellular processes in filamentous fungi

  • The results showed that the canonical G protein-cAMP/PKA pathway plays a pivotal role in the production of chaetoglobosin A (ChA) in C. globosum NK102

  • When the wild type strain was cultured in microcrystalline cellulose (MCC) liquid medium for 4 days and 8 days respectively, we observed that cellulase activity and the cellulase gene expression increased with time in the fungal cultures (Additional file 1: Table S2)

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Summary

Introduction

The canonical heterotrimeric G protein-cAMP/PKA pathway regulates numerous cellular processes in filamentous fungi. Hu et al Microb Cell Fact (2018) 17:160 the reuse of cellulolytic waste [6] Some ascomycetes such as Trichoderma reesei and Aspergillus nidulans are commonly used for producing cellulases and hemicellulases in industry [7, 8]. The cellulose- degrading enzyme system of C. globosum has been studied, and the results suggest an excellent potential for developing a cellulase-producing strain for on-site enzyme production [10]. The transcriptional regulation mechanism of cellulase production in C. globosum is still not clear. A preliminary analysis of putative homologs of the TFs and the distribution of consensus sequences suggests that C. globosum may have a peculiar regulatory mechanism for genes involved in cellulose degradation [19]. The mechanism of cellulase production in C. globosum and the pathways that transduce signals to specific TFs are far unclear

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