Abstract

BackgroundCell wall integrity, vesicle transport and protein secretion are key factors contributing to the vitality and productivity of filamentous fungal cell factories such as Aspergillus niger. In order to pioneer rational strain improvement programs, fundamental knowledge on the genetic basis of these processes is required. The aim of the present study was thus to unravel survival strategies of A. niger when challenged with compounds interfering directly or indirectly with its cell wall integrity: calcofluor white, caspofungin, aureobasidin A, FK506 and fenpropimorph.ResultsTranscriptomics signatures of A. niger and phenotypic analyses of selected null mutant strains were used to predict regulator proteins mediating the survival responses against these stressors. This integrated approach allowed us to reconstruct a model for the cell wall salvage gene network of A. niger that ensures survival of the fungus upon cell surface stress. The model predicts that (i) caspofungin and aureobasidin A induce the cell wall integrity pathway as a main compensatory response via induction of RhoB and RhoD, respectively, eventually activating the mitogen-activated protein kinase kinase MkkA and the transcription factor RlmA. (ii) RlmA is the main transcription factor required for the protection against calcofluor white but it cooperates with MsnA and CrzA to ensure survival of A. niger when challenged with caspofungin and aureobasidin A. (iii) Membrane stress provoked by aureobasidin A via disturbance of sphingolipid synthesis induces cell wall stress, whereas fenpropimorph-induced disturbance of ergosterol synthesis does not.ConclusionThe present work uncovered a sophisticated defence system of A. niger which employs at least three transcription factors - RlmA, MsnA and CrzA – to protect itself against cell wall stress. The transcriptomic data furthermore predicts a fourth transfactor, SrbA, which seems to be specifically important to survive fenpropimorph-induced cell membrane stress. Future studies will disclose how these regulators are interlocked in different signaling pathways to secure survival of A. niger under different cell wall stress conditions.Electronic supplementary materialThe online version of this article (doi:10.1186/s40694-014-0005-8) contains supplementary material, which is available to authorized users.

Highlights

  • Cell wall integrity, vesicle transport and protein secretion are key factors contributing to the vitality and productivity of filamentous fungal cell factories such as Aspergillus niger

  • At least three signaling pathways have been shown to be involved in the cell wall compensatory response in the yeast model Saccharomyces cerevisiae when confronted with cell wall disturbing compounds: the Pkc1p-Slt2p signaling pathway ( named cell wall integrity (CWI) pathway) mediated by the transcription factors Rlm1p and Swi4p/Swi6p, the general stress response pathway mediated by Msn2p/Msn4p, and the calcium/calcineurin pathway mediated by Crz1p [7,8]

  • Spores of the A. niger wild-type strain N402 were allowed to germinate after which they were treated with different concentrations of the sphingolipid biosynthesis inhibitor aureobasidin A (AbaA) and the inhibitor of calcium/calcineurin signaling FK506

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Summary

Introduction

Vesicle transport and protein secretion are key factors contributing to the vitality and productivity of filamentous fungal cell factories such as Aspergillus niger. Fungi are among the most serious biological threats to humans and plants. A preferred target for new antifungal drugs is the fungal cell wall as its composition is fundamentally different from bacterial and plant cell walls. At least three signaling pathways have been shown to be involved in the cell wall compensatory response in the yeast model Saccharomyces cerevisiae when confronted with cell wall disturbing compounds: the Pkc1p-Slt2p signaling pathway ( named cell wall integrity (CWI) pathway) mediated by the transcription factors Rlm1p and Swi4p/Swi6p, the general stress response pathway mediated by Msn2p/Msn4p, and the calcium/calcineurin pathway mediated by Crz1p [7,8]. Whereas cell wall stress responses are well studied and understood in S. cerevisiae, much less is known from Aspergilli, a genus comprising many human and plant pathogenic filamentous fungi. Components of the CWI signaling cascade, the general stress response pathway and the calcium/calcineurin pathway are, conserved among Aspergilli [9,10,11,12,13]

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