Presentation Title

Generation of a Fluorogen-Activating Tag for Candida Albicans Protein Localization

Presenter Information

Christopher DziatkowskiFollow

Advisor Information

Jill Blankenship

Location

Dr. C.C. and Mabel L. Criss Library

Presentation Type

Poster

Start Date

3-3-2017 10:45 AM

End Date

3-3-2017 12:00 PM

Abstract

Candida albicans is a pathogenic fungus that resides in the human body. Normally C. albicans is benign within the human body, however in immunocompromised patients it can pose a significant threat to human health. The mortality rate of such an infection can reach up to 40% even in the event of treatment. Our lab studies a group of proteins, septins, that are imperative for morphogenic conversions that play a key role in the pathogenesis of C. albicans. Furthermore, these septin protein complexes also play a role in antifungal drug susceptibility. Septin localization and abundance within the cell appears to play a role in its response to antifungal drugs and in morphogenic transitions. The use of GFP-tagged septins has revealed novel responses to environmental conditions linked with pathogenesis and C. albicans response to antifungal drugs, however bleaching of the signal means that it is not possible to track individual cell response to these conditions over time. In order to tackle this problem, we are generating a Candida-specific SCFV tag for septin proteins. This antibody-like tag binds to malachite green, this constraining of malachite green creates fluorescence in the green spectrum. This eliminates the bleaching of the signal because of the kinetics of binding between the SCFV and malachite green. This lets us observe septins during stress, giving us insight on their dynamic response to environmental signals. Furthermore, this work will allow us to create a novel tag for C. albicans that could be used to visualize the localization of other stable proteins in the cell.

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COinS
 
Mar 3rd, 10:45 AM Mar 3rd, 12:00 PM

Generation of a Fluorogen-Activating Tag for Candida Albicans Protein Localization

Dr. C.C. and Mabel L. Criss Library

Candida albicans is a pathogenic fungus that resides in the human body. Normally C. albicans is benign within the human body, however in immunocompromised patients it can pose a significant threat to human health. The mortality rate of such an infection can reach up to 40% even in the event of treatment. Our lab studies a group of proteins, septins, that are imperative for morphogenic conversions that play a key role in the pathogenesis of C. albicans. Furthermore, these septin protein complexes also play a role in antifungal drug susceptibility. Septin localization and abundance within the cell appears to play a role in its response to antifungal drugs and in morphogenic transitions. The use of GFP-tagged septins has revealed novel responses to environmental conditions linked with pathogenesis and C. albicans response to antifungal drugs, however bleaching of the signal means that it is not possible to track individual cell response to these conditions over time. In order to tackle this problem, we are generating a Candida-specific SCFV tag for septin proteins. This antibody-like tag binds to malachite green, this constraining of malachite green creates fluorescence in the green spectrum. This eliminates the bleaching of the signal because of the kinetics of binding between the SCFV and malachite green. This lets us observe septins during stress, giving us insight on their dynamic response to environmental signals. Furthermore, this work will allow us to create a novel tag for C. albicans that could be used to visualize the localization of other stable proteins in the cell.