Identification of possible targets of the <it>Aspergillus fumigatus CRZ1 </it>homologue, CrzA

<p>Abstract</p> <p>Background</p> <p>Calcineurin, a serine/threonine-specific protein phosphatase, plays an important role in the control of cell morphology and virulence in fungi. Calcineurin regulates localization and activity of a transcription factor called CRZ1. Re...

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Main Authors: Goldman Gustavo H, Goldman Maria Helena S, Ferreira Márcia ES, Bernardes Luciano AS, Espeso Eduardo, Dinamarco Taísa M, Savoldi Marcela, Malavazi Iran, Soriani Frederico M
Format: Article
Language:English
Published: BMC 2010-01-01
Series:BMC Microbiology
Online Access:http://www.biomedcentral.com/1471-2180/10/12
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author Goldman Gustavo H
Goldman Maria Helena S
Ferreira Márcia ES
Bernardes Luciano AS
Espeso Eduardo
Dinamarco Taísa M
Savoldi Marcela
Malavazi Iran
Soriani Frederico M
author_facet Goldman Gustavo H
Goldman Maria Helena S
Ferreira Márcia ES
Bernardes Luciano AS
Espeso Eduardo
Dinamarco Taísa M
Savoldi Marcela
Malavazi Iran
Soriani Frederico M
author_sort Goldman Gustavo H
collection DOAJ
description <p>Abstract</p> <p>Background</p> <p>Calcineurin, a serine/threonine-specific protein phosphatase, plays an important role in the control of cell morphology and virulence in fungi. Calcineurin regulates localization and activity of a transcription factor called CRZ1. Recently, we characterize <it>Aspergillus fumigatus CRZ1 </it>homologue, AfCrzA. Here, we investigate which pathways are influenced by <it>A. fumigatus </it>AfCrzA during a short pulse of calcium by comparatively determining the transcriptional profile of <it>A. fumigatus </it>wild type and <it>ΔAfcrzA </it>mutant strains.</p> <p>Results</p> <p>We were able to observe 3,622 genes modulated in at least one timepoint in the mutant when compared to the wild type strain (3,211 and 411 at 10 and 30 minutes, respectively). Decreased mRNA abundance in the <it>ΔcrzA </it>was seen for genes encoding calcium transporters, transcription factors and genes that could be directly or indirectly involved in calcium metabolism. Increased mRNA accumulation was observed for some genes encoding proteins involved in stress response. AfCrzA overexpression in <it>A. fumigatus </it>increases the expression of several of these genes. The deleted strain of one of these genes, AfRcnA, belonging to a class of endogenous calcineurin regulators, calcipressins, had more calcineurin activity after exposure to calcium and was less sensitive to menadione 30 μM, hydrogen peroxide 2.5 mM, EGTA 25 mM, and MnCl<sub>2 </sub>25 mM. We constructed deletion, overexpression, and GFP fusion protein for the closely related <it>A. nidulans </it>AnRcnA. GFP::RcnA was mostly detected along the germling, did not accumulate in the nuclei and its location is not affected by the cellular response to calcium chloride.</p> <p>Conclusion</p> <p>We have performed a transcriptional profiling analysis of the <it>A. fumigatus ΔAfcrzA </it>mutant strain exposed to calcium stress. This provided an excellent opportunity to identify genes and pathways that are under the influence of AfCrzA. AfRcnA, one of these selected genes, encodes a modulator of calcineurin activity. Concomitantly with <it>A. fumigatus AfrcnA </it>molecular analysis, we decided to exploit the conserved features of <it>A. nidulans </it>calcineurin system and investigated the <it>A. nidulans </it>AnRcnA homologue. <it>A. nidulans </it>AnRcnA mutation is suppressing CnaA mutation and it is responsible for modulating the calcineurin activity and mRNA accumulation of genes encoding calcium transporters.</p>
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spelling doaj.art-1bf6ff3fd6264e719ebc5931226252c22022-12-22T03:26:13ZengBMCBMC Microbiology1471-21802010-01-011011210.1186/1471-2180-10-12Identification of possible targets of the <it>Aspergillus fumigatus CRZ1 </it>homologue, CrzAGoldman Gustavo HGoldman Maria Helena SFerreira Márcia ESBernardes Luciano ASEspeso EduardoDinamarco Taísa MSavoldi MarcelaMalavazi IranSoriani Frederico M<p>Abstract</p> <p>Background</p> <p>Calcineurin, a serine/threonine-specific protein phosphatase, plays an important role in the control of cell morphology and virulence in fungi. Calcineurin regulates localization and activity of a transcription factor called CRZ1. Recently, we characterize <it>Aspergillus fumigatus CRZ1 </it>homologue, AfCrzA. Here, we investigate which pathways are influenced by <it>A. fumigatus </it>AfCrzA during a short pulse of calcium by comparatively determining the transcriptional profile of <it>A. fumigatus </it>wild type and <it>ΔAfcrzA </it>mutant strains.</p> <p>Results</p> <p>We were able to observe 3,622 genes modulated in at least one timepoint in the mutant when compared to the wild type strain (3,211 and 411 at 10 and 30 minutes, respectively). Decreased mRNA abundance in the <it>ΔcrzA </it>was seen for genes encoding calcium transporters, transcription factors and genes that could be directly or indirectly involved in calcium metabolism. Increased mRNA accumulation was observed for some genes encoding proteins involved in stress response. AfCrzA overexpression in <it>A. fumigatus </it>increases the expression of several of these genes. The deleted strain of one of these genes, AfRcnA, belonging to a class of endogenous calcineurin regulators, calcipressins, had more calcineurin activity after exposure to calcium and was less sensitive to menadione 30 μM, hydrogen peroxide 2.5 mM, EGTA 25 mM, and MnCl<sub>2 </sub>25 mM. We constructed deletion, overexpression, and GFP fusion protein for the closely related <it>A. nidulans </it>AnRcnA. GFP::RcnA was mostly detected along the germling, did not accumulate in the nuclei and its location is not affected by the cellular response to calcium chloride.</p> <p>Conclusion</p> <p>We have performed a transcriptional profiling analysis of the <it>A. fumigatus ΔAfcrzA </it>mutant strain exposed to calcium stress. This provided an excellent opportunity to identify genes and pathways that are under the influence of AfCrzA. AfRcnA, one of these selected genes, encodes a modulator of calcineurin activity. Concomitantly with <it>A. fumigatus AfrcnA </it>molecular analysis, we decided to exploit the conserved features of <it>A. nidulans </it>calcineurin system and investigated the <it>A. nidulans </it>AnRcnA homologue. <it>A. nidulans </it>AnRcnA mutation is suppressing CnaA mutation and it is responsible for modulating the calcineurin activity and mRNA accumulation of genes encoding calcium transporters.</p>http://www.biomedcentral.com/1471-2180/10/12
spellingShingle Goldman Gustavo H
Goldman Maria Helena S
Ferreira Márcia ES
Bernardes Luciano AS
Espeso Eduardo
Dinamarco Taísa M
Savoldi Marcela
Malavazi Iran
Soriani Frederico M
Identification of possible targets of the <it>Aspergillus fumigatus CRZ1 </it>homologue, CrzA
BMC Microbiology
title Identification of possible targets of the <it>Aspergillus fumigatus CRZ1 </it>homologue, CrzA
title_full Identification of possible targets of the <it>Aspergillus fumigatus CRZ1 </it>homologue, CrzA
title_fullStr Identification of possible targets of the <it>Aspergillus fumigatus CRZ1 </it>homologue, CrzA
title_full_unstemmed Identification of possible targets of the <it>Aspergillus fumigatus CRZ1 </it>homologue, CrzA
title_short Identification of possible targets of the <it>Aspergillus fumigatus CRZ1 </it>homologue, CrzA
title_sort identification of possible targets of the it aspergillus fumigatus crz1 it homologue crza
url http://www.biomedcentral.com/1471-2180/10/12
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