Event Title

Genetic Analysis of Interacting Proteins

Presenter Information

Noah Sutton, Minnesota State University, Mankato

Location

CSU 255

Start Date

25-4-2005 1:15 PM

End Date

25-4-2005 3:15 PM

Student's Major

Biological Sciences

Student's College

Science, Engineering and Technology

Mentor's Name

Marilyn Hart

Mentor's Department

Biological Sciences

Mentor's College

Social and Behavioral Sciences

Description

Actin, a component of all eukaryotic cells, contributes to cell motility and shape. Actin capping protein (CP), associated with the actin cytoskeleton, is a heterodimer composed of alpha and beta subunits. The beta subunit has three isoforms: beta l (β1), beta 2 (β2), and beta 3 (β3). These isoforms are produced from alternative splicing of one gene with 90% sequence identity. The region of divergence defines membership to each subfamily. Previous work in cardiac myocytes has shown that β1 cannot functionally replace β2 nor can β2 functionally replace β1. Furthermore, data suggests that the isoform specific functions are due to novel protein interactions. We have identified proteins that interact with each isoform via a yeast two-hybrid genetic screen. The screen utilized two specific components: a bait plasmid and a prey plasmid. We have generated bait constructs, confirmed their orientation and expression, and executed a large-scale genetic screen. 14 β1 and 213 β2 interacting constructs have been identified. The genetic interactions have been confirmed using both expression of beta-galactosidase and histidine synthesis. Additional preliminary characterization of the constructs will be presented.