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Transcriptional profile of Paracoccidioides spp. in response to itraconazole.

da Silva Neto BR, Carvalho PF, Bailão AM, Martins WS, Soares CM, Pereira M - BMC Genomics (2014)

Bottom Line: Among the genes affected, we identified genes in common with other fungi, as well as genes unique to Paracoccidioides Pb01.Voltage-gated Ca2+ alpha subunit (CAV), Tetracycline resistance protein (TETA) and Hemolisyn-iii channel protein (HLYiii) were found only here and a probably involvement with resistance to itraconazole could be investigated in the future.However our findings do not permit inference to current clinical practice.

View Article: PubMed Central - HTML - PubMed

Affiliation: Departamento de Bioquímica e Biologia Molecular, Laboratório de Biologia Molecular, Instituto de Ciências Biológicas, ICBII, Campus II, Universidade Federal de Goiás, C,P, 131, 74001-970 Goiânia, GO, Brazil. maristelaufg@gmail.com.

ABSTRACT

Background: Itraconazole is currently used to treat paracoccidioidomycosis. The mechanism of action of azoles has been elucidated in some fungi, although little is known regarding its mechanism of action in Paracoccidioides spp. The present work focused on identification of regulated transcripts using representational difference analysis of Paracoccidioides spp. yeast cells treated with itraconazole for 1 and 2 h.

Results: Paracoccidioides Pb01 genes up-regulated by itraconazole included genes involved in cellular transport, metabolism/energy, transcription, cell rescue, defense and virulence. ERG11, ERG6, ERG3, ERG5 and ERG25 were up-regulated at multiple time points. In vivo infection experiments in mice corroborated the in vitro results. Ergosterol levels and distribution were evaluated in Paracoccidioides Pb18 yeast cells, and the results demonstrate that both factors were changed in the fungus treated with itraconazole.

Conclusion: To our knowledge, this is the first transcriptional analysis of Paracoccidioides spp. exposed to a triazole drug. Here acetyl seems to be intensively produced from different metabolic pathways to produce ergosterol by the action of ergosterol synthesis related enzymes, which were also affected in other fungi. Among the genes affected, we identified genes in common with other fungi, as well as genes unique to Paracoccidioides Pb01. Those genes could be considered target to new drugs. Voltage-gated Ca2+ alpha subunit (CAV), Tetracycline resistance protein (TETA) and Hemolisyn-iii channel protein (HLYiii) were found only here and a probably involvement with resistance to itraconazole could be investigated in the future. However our findings do not permit inference to current clinical practice.

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Distribution of genes responding to itraconazole in P. brasiliensis isolate Pb 01. Data are shown for a subset of genes that were significantly up or down regulated (e-values ≤10-10). The search for functional categories was performed by using the Blast2GO program that joints in one application GO annotation based on similarity searches with statistical analysis and highlight visualization on directed acyclic graphs. GO terms shown are those that were considered significantly over represented by the analysis. Sequences were grouped in functional categories according to the classification of the MIPS functional catalog (Munich Center for Protein Sequences; http://mips.gsf.de/). Specific genes for P. brasiliensis isolate Pb01 are underlined, genes found in other fungi when exposed to itraconazole and other azoles are represented with * and represented with ◊, respectively. Numbers in parentheses represent changes in gene expression. Positive signal indicate induction, and negative indicate repression.
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Figure 5: Distribution of genes responding to itraconazole in P. brasiliensis isolate Pb 01. Data are shown for a subset of genes that were significantly up or down regulated (e-values ≤10-10). The search for functional categories was performed by using the Blast2GO program that joints in one application GO annotation based on similarity searches with statistical analysis and highlight visualization on directed acyclic graphs. GO terms shown are those that were considered significantly over represented by the analysis. Sequences were grouped in functional categories according to the classification of the MIPS functional catalog (Munich Center for Protein Sequences; http://mips.gsf.de/). Specific genes for P. brasiliensis isolate Pb01 are underlined, genes found in other fungi when exposed to itraconazole and other azoles are represented with * and represented with ◊, respectively. Numbers in parentheses represent changes in gene expression. Positive signal indicate induction, and negative indicate repression.

Mentions: Among the Paracoccidioides Pb01 genes regulated by itraconazole were those involved in cellular transport, metabolism/energy, transcription, cell rescue, defense and virulence. Similar and different groups were also observed in other fungi in response to different azoles [8,9,11,12,15] (Figure 5). Among the genes affected, we identified genes in common with other fungi, as well as genes unique to Paracoccidioides. In fact few Paracoccidioides spp. genes were shared with genes from other fungi. This could be due to different techniques and classes of azoles used in the works. The comparison with ither fungi show that cell processes related to stress response, xenobiotic efflux are trigered upon itraconazol in different fungi. The genes exclusively regulated in Paracoccidioides spp. reveal that fungi response to drugs can partially involve specific processes that may be related to different sensibility of differetn fungi to itraconazol treatment. This could be due to different techniques and classes of azoles used in the works.


Transcriptional profile of Paracoccidioides spp. in response to itraconazole.

da Silva Neto BR, Carvalho PF, Bailão AM, Martins WS, Soares CM, Pereira M - BMC Genomics (2014)

Distribution of genes responding to itraconazole in P. brasiliensis isolate Pb 01. Data are shown for a subset of genes that were significantly up or down regulated (e-values ≤10-10). The search for functional categories was performed by using the Blast2GO program that joints in one application GO annotation based on similarity searches with statistical analysis and highlight visualization on directed acyclic graphs. GO terms shown are those that were considered significantly over represented by the analysis. Sequences were grouped in functional categories according to the classification of the MIPS functional catalog (Munich Center for Protein Sequences; http://mips.gsf.de/). Specific genes for P. brasiliensis isolate Pb01 are underlined, genes found in other fungi when exposed to itraconazole and other azoles are represented with * and represented with ◊, respectively. Numbers in parentheses represent changes in gene expression. Positive signal indicate induction, and negative indicate repression.
© Copyright Policy - open-access
Related In: Results  -  Collection

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Show All Figures
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Figure 5: Distribution of genes responding to itraconazole in P. brasiliensis isolate Pb 01. Data are shown for a subset of genes that were significantly up or down regulated (e-values ≤10-10). The search for functional categories was performed by using the Blast2GO program that joints in one application GO annotation based on similarity searches with statistical analysis and highlight visualization on directed acyclic graphs. GO terms shown are those that were considered significantly over represented by the analysis. Sequences were grouped in functional categories according to the classification of the MIPS functional catalog (Munich Center for Protein Sequences; http://mips.gsf.de/). Specific genes for P. brasiliensis isolate Pb01 are underlined, genes found in other fungi when exposed to itraconazole and other azoles are represented with * and represented with ◊, respectively. Numbers in parentheses represent changes in gene expression. Positive signal indicate induction, and negative indicate repression.
Mentions: Among the Paracoccidioides Pb01 genes regulated by itraconazole were those involved in cellular transport, metabolism/energy, transcription, cell rescue, defense and virulence. Similar and different groups were also observed in other fungi in response to different azoles [8,9,11,12,15] (Figure 5). Among the genes affected, we identified genes in common with other fungi, as well as genes unique to Paracoccidioides. In fact few Paracoccidioides spp. genes were shared with genes from other fungi. This could be due to different techniques and classes of azoles used in the works. The comparison with ither fungi show that cell processes related to stress response, xenobiotic efflux are trigered upon itraconazol in different fungi. The genes exclusively regulated in Paracoccidioides spp. reveal that fungi response to drugs can partially involve specific processes that may be related to different sensibility of differetn fungi to itraconazol treatment. This could be due to different techniques and classes of azoles used in the works.

Bottom Line: Among the genes affected, we identified genes in common with other fungi, as well as genes unique to Paracoccidioides Pb01.Voltage-gated Ca2+ alpha subunit (CAV), Tetracycline resistance protein (TETA) and Hemolisyn-iii channel protein (HLYiii) were found only here and a probably involvement with resistance to itraconazole could be investigated in the future.However our findings do not permit inference to current clinical practice.

View Article: PubMed Central - HTML - PubMed

Affiliation: Departamento de Bioquímica e Biologia Molecular, Laboratório de Biologia Molecular, Instituto de Ciências Biológicas, ICBII, Campus II, Universidade Federal de Goiás, C,P, 131, 74001-970 Goiânia, GO, Brazil. maristelaufg@gmail.com.

ABSTRACT

Background: Itraconazole is currently used to treat paracoccidioidomycosis. The mechanism of action of azoles has been elucidated in some fungi, although little is known regarding its mechanism of action in Paracoccidioides spp. The present work focused on identification of regulated transcripts using representational difference analysis of Paracoccidioides spp. yeast cells treated with itraconazole for 1 and 2 h.

Results: Paracoccidioides Pb01 genes up-regulated by itraconazole included genes involved in cellular transport, metabolism/energy, transcription, cell rescue, defense and virulence. ERG11, ERG6, ERG3, ERG5 and ERG25 were up-regulated at multiple time points. In vivo infection experiments in mice corroborated the in vitro results. Ergosterol levels and distribution were evaluated in Paracoccidioides Pb18 yeast cells, and the results demonstrate that both factors were changed in the fungus treated with itraconazole.

Conclusion: To our knowledge, this is the first transcriptional analysis of Paracoccidioides spp. exposed to a triazole drug. Here acetyl seems to be intensively produced from different metabolic pathways to produce ergosterol by the action of ergosterol synthesis related enzymes, which were also affected in other fungi. Among the genes affected, we identified genes in common with other fungi, as well as genes unique to Paracoccidioides Pb01. Those genes could be considered target to new drugs. Voltage-gated Ca2+ alpha subunit (CAV), Tetracycline resistance protein (TETA) and Hemolisyn-iii channel protein (HLYiii) were found only here and a probably involvement with resistance to itraconazole could be investigated in the future. However our findings do not permit inference to current clinical practice.

Show MeSH
Related in: MedlinePlus