Analysis of Bm-daf-16a Function in Brugia malayi
Mentor 1
Kirsten Crossgrove
Location
Union Wisconsin Room
Start Date
24-4-2015 2:30 PM
End Date
24-4-2015 3:45 PM
Description
We hypothesize that the Bm-daf-16a gene plays an important role in the developmental transition of the infective L3 stage to L4 of the parasitic nematode Brugia malayi. DAF-16 is a transcriptional regulator that is modulated by the insulin/IGF-1 signaling (IIS) pathway. In the model organism Caenorhabditis elegans, DAF-16 is required for dauer formation, an alternate stage of development induced by stressful conditions. Dauer larvae of C. elegans are analogous to L3 infective larvae of B. malayi, suggesting that Bm-DAF-16 is important for development and infection of the parasite. To test whether Bm-DAF-16 is regulated by insulin signaling, we cloned Bm-daf-16a into a mammalian expression vector. We are transfecting mammalian cells with Bm-daf-16a and treating the cells to both high and low levels of serum (contains insulin). We predict that transfected cells treated with low serum levels (low insulin signaling) will report higher Bm-DAF-16 activity. We will monitor Bm-DAF-16 activity using luciferase reporter genes with DAF-16 response elements upstream. If Bm-daf-16a is shown to be negatively regulated by insulin signaling, we will try to find its subcellular location by tagging the expressed DAF-16 protein with fluorescent ligands that can be visualized under a fluorescent microscope. We expect to find DAF-16 in the cytoplasm in the high serum levels and in the nucleus when the cells are put in low serum levels. These findings will lead to a greater understanding of the filarial nematode life cycle and could lead to new treatment options for people infected with Brugia malayi.
Analysis of Bm-daf-16a Function in Brugia malayi
Union Wisconsin Room
We hypothesize that the Bm-daf-16a gene plays an important role in the developmental transition of the infective L3 stage to L4 of the parasitic nematode Brugia malayi. DAF-16 is a transcriptional regulator that is modulated by the insulin/IGF-1 signaling (IIS) pathway. In the model organism Caenorhabditis elegans, DAF-16 is required for dauer formation, an alternate stage of development induced by stressful conditions. Dauer larvae of C. elegans are analogous to L3 infective larvae of B. malayi, suggesting that Bm-DAF-16 is important for development and infection of the parasite. To test whether Bm-DAF-16 is regulated by insulin signaling, we cloned Bm-daf-16a into a mammalian expression vector. We are transfecting mammalian cells with Bm-daf-16a and treating the cells to both high and low levels of serum (contains insulin). We predict that transfected cells treated with low serum levels (low insulin signaling) will report higher Bm-DAF-16 activity. We will monitor Bm-DAF-16 activity using luciferase reporter genes with DAF-16 response elements upstream. If Bm-daf-16a is shown to be negatively regulated by insulin signaling, we will try to find its subcellular location by tagging the expressed DAF-16 protein with fluorescent ligands that can be visualized under a fluorescent microscope. We expect to find DAF-16 in the cytoplasm in the high serum levels and in the nucleus when the cells are put in low serum levels. These findings will lead to a greater understanding of the filarial nematode life cycle and could lead to new treatment options for people infected with Brugia malayi.
