Evaluating the role of the tandem repeat region in the Epa1p fungal adhesin of Candida glabrata
Raposo, Colin J.
2019
- Infection by Candida glabrata is reliant on a suite of cell surface adhesins that play a variety of roles necessary for transmission, establishment, and proliferation of C. glabrata infection. One particular adhesin, Epithelial Adhesin 1 (Epa1p), is responsible for binding to host tissue and biofilm formation, which is essential for fungal propagation. Epa1p structure consists of three domains: ... read morea C-terminal GPI anchor for cell wall linkage, an N-terminal intercellular binding domain responsible for epithelial cell binding, and a Serine / Threonine-rich linker domain connecting these terminal domains. The linker domain contains a 40-amino acid tandem repeat region which our lab has observed to be variable in repeat copy number between isolates from clinical sources. Based om the relationship between repeat copy number and binding capacity of other adhesins with similar structure, we hypothesized that Epa1p repeat copy number may encode variable protein function. To test this, we expressed Epa1p with various repeat copy numbers in S. cerevisiae to determine how differences in repeat copy number affect Epa1p expression, surface display and binding to human epithelial cells. Our data suggest that expression and display of Epa1p improves with decreased repeat copy number. We also have collected data, which show that Epa1p constructs with a deleted repeat region or a lower repeat copy number show lower levels of stability in the cell wall and decreased ability to bind to cells at the single protein level. Together these data show that repeat copy number of Epa1p influences the properties of Epa1p and the adhesive characteristics of C. glabrata.read less
- ID:
- 9306tb29h
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