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Time-course proteome analysis reveals the dynamic response of Cryptococcus gattii cells to fluconazole.

Chong HS, Campbell L, Padula MP, Hill C, Harry E, Li SS, Wilkins MR, Herbert B, Carter D - PLoS ONE (2012)

Bottom Line: During FLC treatment, an increase in stress response, ATP synthesis and mitochondrial respiratory chain proteins, and a decrease in most ribosomal proteins was observed, suggesting that ATP-dependent efflux pumps had been initiated for survival and that the maintenance of ribosome synthesis was differentially expressed.An integrative network analysis revealed co-ordinated processes involved in drug response, and highlighted hubs in the network representing essential proteins that are required for cell viability.This work demonstrates the dynamic cellular response of a typical susceptible isolate of C. gattii to FLC, and identified a number of proteins and pathways that could be targeted to augment the activity of FLC.

View Article: PubMed Central - PubMed

Affiliation: School of Molecular Bioscience, University of Sydney, Sydney, New South Wales, Australia.

ABSTRACT
Cryptococcus gattii is an encapsulated fungus capable of causing fatal disease in immunocompetent humans and animals. As current antifungal therapies are few and limited in efficacy, and resistance is an emerging issue, the development of new treatment strategies is urgently required. The current study undertook a time-course analysis of the proteome of C. gattii during treatment with fluconazole (FLC), which is used widely in prophylactic and maintenance therapies. The aims were to analyze the overall cellular response to FLC, and to find fungal proteins involved in this response that might be useful targets in therapies that augment the antifungal activity of FLC. During FLC treatment, an increase in stress response, ATP synthesis and mitochondrial respiratory chain proteins, and a decrease in most ribosomal proteins was observed, suggesting that ATP-dependent efflux pumps had been initiated for survival and that the maintenance of ribosome synthesis was differentially expressed. Two proteins involved in fungal specific pathways were responsive to FLC. An integrative network analysis revealed co-ordinated processes involved in drug response, and highlighted hubs in the network representing essential proteins that are required for cell viability. This work demonstrates the dynamic cellular response of a typical susceptible isolate of C. gattii to FLC, and identified a number of proteins and pathways that could be targeted to augment the activity of FLC.

Show MeSH
The effect of FLC treatment on the morphology of C. gattii cells.Data are shown as mean +/− standard error. * indicates P<0.05 and ** indicates P<0.001.
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pone-0042835-g002: The effect of FLC treatment on the morphology of C. gattii cells.Data are shown as mean +/− standard error. * indicates P<0.05 and ** indicates P<0.001.

Mentions: Growth of a representative, moderately susceptible C. gattii isolate was initially assessed in the presence of 20 and 100 µg/mL FLC. Growth of all cell cultures was affected after exposure to FLC for 2 hours (Figure 1), and cell imaging revealed changes to cellular morphology during FLC treatment (Figure 2). There were no significant differences in capsule size among cells treated with or without FLC at each time point (p≥0.65). However, cells treated with either concentration of FLC for 4 h and 6 h were significantly larger than the untreated cells (p<0.001) and the cells treated with FLC for 3 h (p≤0.004). Cells treated with 20 µg/mL of FLC had a similar size to cells treated with 100 µg/mL of FLC at three different time points (p≥0.25). There was no difference in the size of cells treated with FLC for 4 h and 6 h (p≥0.15), or between untreated cells and those treated with FLC for 3 h (p≥0.07).


Time-course proteome analysis reveals the dynamic response of Cryptococcus gattii cells to fluconazole.

Chong HS, Campbell L, Padula MP, Hill C, Harry E, Li SS, Wilkins MR, Herbert B, Carter D - PLoS ONE (2012)

The effect of FLC treatment on the morphology of C. gattii cells.Data are shown as mean +/− standard error. * indicates P<0.05 and ** indicates P<0.001.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3412811&req=5

pone-0042835-g002: The effect of FLC treatment on the morphology of C. gattii cells.Data are shown as mean +/− standard error. * indicates P<0.05 and ** indicates P<0.001.
Mentions: Growth of a representative, moderately susceptible C. gattii isolate was initially assessed in the presence of 20 and 100 µg/mL FLC. Growth of all cell cultures was affected after exposure to FLC for 2 hours (Figure 1), and cell imaging revealed changes to cellular morphology during FLC treatment (Figure 2). There were no significant differences in capsule size among cells treated with or without FLC at each time point (p≥0.65). However, cells treated with either concentration of FLC for 4 h and 6 h were significantly larger than the untreated cells (p<0.001) and the cells treated with FLC for 3 h (p≤0.004). Cells treated with 20 µg/mL of FLC had a similar size to cells treated with 100 µg/mL of FLC at three different time points (p≥0.25). There was no difference in the size of cells treated with FLC for 4 h and 6 h (p≥0.15), or between untreated cells and those treated with FLC for 3 h (p≥0.07).

Bottom Line: During FLC treatment, an increase in stress response, ATP synthesis and mitochondrial respiratory chain proteins, and a decrease in most ribosomal proteins was observed, suggesting that ATP-dependent efflux pumps had been initiated for survival and that the maintenance of ribosome synthesis was differentially expressed.An integrative network analysis revealed co-ordinated processes involved in drug response, and highlighted hubs in the network representing essential proteins that are required for cell viability.This work demonstrates the dynamic cellular response of a typical susceptible isolate of C. gattii to FLC, and identified a number of proteins and pathways that could be targeted to augment the activity of FLC.

View Article: PubMed Central - PubMed

Affiliation: School of Molecular Bioscience, University of Sydney, Sydney, New South Wales, Australia.

ABSTRACT
Cryptococcus gattii is an encapsulated fungus capable of causing fatal disease in immunocompetent humans and animals. As current antifungal therapies are few and limited in efficacy, and resistance is an emerging issue, the development of new treatment strategies is urgently required. The current study undertook a time-course analysis of the proteome of C. gattii during treatment with fluconazole (FLC), which is used widely in prophylactic and maintenance therapies. The aims were to analyze the overall cellular response to FLC, and to find fungal proteins involved in this response that might be useful targets in therapies that augment the antifungal activity of FLC. During FLC treatment, an increase in stress response, ATP synthesis and mitochondrial respiratory chain proteins, and a decrease in most ribosomal proteins was observed, suggesting that ATP-dependent efflux pumps had been initiated for survival and that the maintenance of ribosome synthesis was differentially expressed. Two proteins involved in fungal specific pathways were responsive to FLC. An integrative network analysis revealed co-ordinated processes involved in drug response, and highlighted hubs in the network representing essential proteins that are required for cell viability. This work demonstrates the dynamic cellular response of a typical susceptible isolate of C. gattii to FLC, and identified a number of proteins and pathways that could be targeted to augment the activity of FLC.

Show MeSH