Genetic Analysis of Interacting Proteins
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.
Genetic Analysis of Interacting Proteins
CSU 255
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.
Recommended Citation
Sutton, Noah. "Genetic Analysis of Interacting Proteins." Undergraduate Research Symposium, Mankato, MN, April 25, 2005.
https://cornerstone.lib.mnsu.edu/urs/2005/poster-session-B/4
