Study of cell wall integrity and disturbing antifungal compounds with a novel GFP-based reporter method.

Robert Damveld 1, John van Dam 1, Frans Klis 2, C.A.M.J.J. van den Hondel1,3 and A.F.J. Ram

Author address: 

Institute of Biology,W assenaarseweg 64, 2333 AL Leiden, The Netherlands, 2 Swammerdaam Institute for Life Sciences, University of Amsterdam, Nieuwe Achtergracht 166, 1018 W V Amsterdam 3Department of Applied Microbiology and Gene Technology, TNO-

Abstract: 

Activation of the fungal cell wall integrity pathway is a mechanism used by fungi to escape from cell wall threatening conditions. In Aspergillus niger we have previously shown that the agsA gene, encoding an í¡-1,3-glucan synthase submit is transcriptionally activated in response to cell wall stress, and that this response is mediated via a RLM A dependent signal transduction pathway. W e have used the agsA promoter region to set up a GFP-based reporter system to identify compounds that activate the agsA expression by fusing the PagsA to the cytosolic GFP or to a nuclear targeted version of GFP. The reporter strains confirmed earlier observation that the agsA gene is activated in response to various cell wall disturbing compounds such as CFW , caspofungin and tunicamycin. Other forms of stress (osmotic or oxidative stress) did not activate agsA expression, indicating that the induction of agsA is cell wall stress specific and not a general stress response. W e have used the cell wall stress reporter system to study the effect of 20 known antifungal compounds in relation to cell wall remodeling.
2005

abstract No: 

63.

Full conference title: 

23rd Fungal Genetics Conference
    • Fungal Genetics Conference 23rd (2002)