Determining binding specificities of cell adhesion molecules from Drosophila
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Date
2016-09-15
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Abstract
Neurons of both vertebrates and invertebrates exhibit a complex set of cell-to-cell interactions during successful development of the nervous system. Cell adhesion molecules (CAMs) play an important role in mediating many of these specific and stereotyped cell-cell interactions. I am investigating the binding specificities of two CAMs from Dipteran insects: Lachesin (Lac) and Amalgam (Ama). Ama arose as a duplication of Lac in early Dipteran evolution, and both proteins still share extensive amino acid similarity. Lac, which is membrane-linked, homophilically binds itself. Ama, secreted into the extracellular matrix, has both a homophilic binding property as well as the ability to heterophilically bind the transmembrane protein Neurotactin (Nrt). Despite the high level of amino acid sequence similarity between Ama and Lac, the two proteins are unable to bind each other, and Lac does not display an interaction with Nrt. Therefore, I am identifying the precise domain(s) of Lac and Ama that produce these differences in binding specificity. To accomplish this, chimeric constructs of the three immunoglobulin-like domains of ama and lac from D. melanogaster have been created and cloned into a vector for regulated expression in Schneider 2 (S2) cells. The S2 cells are then to be used for aggregation assays, which allow us to observe the binding properties of these proteins. In addition, I am cloning the ama and lac orthologues from other Dipteran species to better understand how the unique binding properties of these two proteins have evolved. This project will help to further characterize the complex series of intercellular interactions during nervous system development.
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Biological and Biomedical Sciences
Keywords
cell adhesion molecules, neurogenesis, Drosophila, molecular genetics