title: Modulation of fungal sensitivity to staurosporine by targeting proteins identified by transcriptional profiling.
abstract: An analysis of the time-dependent genetic response to the death-inducer staurosporine was performed in Neurospora crassa by transcriptional profiling. Staurosporine induced two major genes encoding an ABC transporter and a protein with similarity to regulatory subunits of potassium channels. The transcriptional response is dependent on the activity of a novel transcription factor. Deletion mutants in differentially expressed genes displayed altered sensitivity to staurosporine, underscoring significant proteins involved in the response to the drug. A null-mutant of the ABC transporter (abc3) is extremely sensitive to staurosporine, accumulates more staurosporine than the wild type strain and is defective in energy-dependent export of the drug, indicating that the ABC3 protein is the first described staurosporine transporter. It was located in the plasma membrane by immunofluorescence microscopy. The combination of inhibitors of ABC transporters or of potassium channels with staurosporine leads to an enhanced activity against N. crassa and pathogenic fungi paving the way to the development of more potent and specific antifungals. Our results highlight the general use of transcriptional profiling for the identification of novel proteins involved in cell death and their potential use as drug targets.
authors: IBMC - Instituto de Biologia Molecular e Celular, Portugal.
authors: Andreia S
authors: AS
authors: Fernandes
authors: A Pedro
authors: AP
authors: Gonçalves
authors: Ana
authors: A
authors: Castro
authors: Telma A
authors: TA
authors: Lopes
authors: Rui
authors: R
authors: Gardner
authors: N Louise
authors: NL
authors: Glass
authors: Arnaldo
authors: A
authors: Videira
mesh_terms: D015761:4-Aminopyridine / Q000494:pharmacology; D018528:ATP-Binding Cassette Transporters / Q000235:genetics / Q000378:metabolism; D017209:Apoptosis / Q000187:drug effects / Q000235:genetics; D005656:Fungal Proteins / Q000235:genetics / Q000378:metabolism*; D015870:Gene Expression / Q000187:drug effects*; D020869:Gene Expression Profiling; D015966:Gene Expression Regulation, Fungal / Q000187:drug effects; D005800:Genes, Fungal / Q000187:drug effects; D007150:Immunohistochemistry; D020287:In Situ Nick-End Labeling; D046228:Microarray Analysis; D009492:Neurospora crassa / Q000187:drug effects / Q000235:genetics* / Q000378:metabolism*; D019311:Staurosporine / Q000378:metabolism / Q000494:pharmacology*; D014157:Transcription Factors / Q000378:metabolism
chemical_list: D018528:ATP-Binding Cassette Transporters; D005656:Fungal Proteins; D014157:Transcription Factors; D015761:4-Aminopyridine; D019311:Staurosporine