Submission Date

4-30-2020

Document Type

Paper- Restricted to Campus Access

Department

Biology

Adviser

Jennifer Round

Committee Member

Rebecca Lyczak

Committee Member

John Spencer

Department Chair

Rebecca Lyczak

External Reviewer

Radha Aras

Distinguished Honors

This paper has met the requirements for Distinguished Honors.

Project Description

In the developing nervous system, synapse formation is crucial to establish proper brain wiring and therefore function. At synapses, several pre- and postsynaptic adhesion proteins are expressed to coordinate synapse formation. Slitrks are a family of postsynaptic adhesion proteins that contribute to this process. While extracellular binding partners of Slitrks have been identified, little is known about their intracellular binding partners. Our lab recently identified an interaction between Slitrk2 and PSD-95, an intracellular MAGUK scaffold protein. The focus of this research was to map the Slitrk2/PSD-95 interaction using a yeast two-hybrid assay. In order to identify the domains of each protein required for the interaction, full length, truncated, and mutated forms of each protein were fused to transcription factor domains and expressed in yeast. Positive interactions were detected by blue yeast growth on selective media initiated by the transcription of reporter genes. It was determined that the SH3 domain of PSD-95 along with the last four amino acids and an aspartic acid-tyrosine motif of Slitrk2 mediate their interaction. Successful confirmation of protein expression in yeast was achieved through separation of whole cell protein extracts via SDS-PAGE and Western blotting. Identifying the critical domains of interaction between Slitrk2 and PSD-95 will allow us to identify the functional significance of the interaction in neurons. Overall, understanding Slitrks and their intracellular binding partners further contributes to our understanding of synaptic adhesion during nervous system development.

Comments

Project funded by Ursinus College (Ursinus College Summer Fellows Program, the Ursinus Student Achievement in Research and Creativity Committee, and the Ursinus College Biology Department) and The Pittsburgh Foundation.

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