TY - JOUR
T1 - Modulation of fungal sensitivity to staurosporine by targeting proteins identified by transcriptional profiling
AU - Fernandes, Andreia S.
AU - Pedro Gonçalves, A.
AU - Castro, Ana
AU - Lopes, Telma A.
AU - Gardner, Rui
AU - Louise Glass, N.
AU - Videira, Arnaldo
N1 - Funding Information:
We acknowledge Paula Magalhães for help with Real-Time PCR, Dr Isabel Carvalho and Tânia Ribeiro for rat immunization and antiserum preparation, and Dr Paula Sampaio for help with immunofluorescence microscopy. This work was supported by a “Programa Ciência” fellowship financed by POPH-QREN (typology 4.2) with co-funding from ESF and MCTES to ASF, a Fundação Calouste Gulbenkian PhD fellowship to APG, NIH grant GM60468 to NLG, research grants from FCT Portugal and the European POCI program of QCAIII (co-participated by FEDER) and a sabbatical fellowship from Fundação Luso-Americana to AV.
PY - 2011/12
Y1 - 2011/12
N2 - 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.
AB - 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.
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U2 - 10.1016/j.fgb.2011.09.004
DO - 10.1016/j.fgb.2011.09.004
M3 - Article
C2 - 22001288
AN - SCOPUS:82555176625
SN - 1087-1845
VL - 48
SP - 1130
EP - 1138
JO - Fungal Genetics and Biology
JF - Fungal Genetics and Biology
IS - 12
ER -