Identification of Proteins Interacting with the Alpha Subunits of Actin Capping Protein
Location
CSU 203
Start Date
2-4-2019 1:05 PM
End Date
2-4-2019 2:05 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
Science, Engineering and Technology
Description
Capping protein (CP) is an actin binding protein composed of an alpha and a beta subunit, which is important for actin assembly and cell motility. Whereas lower organisms have one gene and one isoform of each subunit, higher organisms have multiple alpha isoforms (a1, a2 and a3) with conserved sequences defining highly conserved subfamilies. CPa1 and CPa2 share >90% sequence identity and the regions of divergence are highly conserved among the subunits across vertebrate. In addition, CPa1 and CPa2have distinct expression patterns in murine tissues. Endothelial cells contain only CPa2, and erythrocytes contain almost exclusively CPa1. Most tissues have both isoforms but the ratio of alpha1:alpha2 varies widely. The highly conserved sequence conservation and distinct expression patterns of CPa1 and CPa2 support our hypothesis that the CP alpha isoforms have conserved, unique and essential roles in vertebrates and will therefore, interact with unique proteins. We executed a yeast two-hybrid screen to identify proteins that interact with CPa1 and CPa2. To date, approximately 100 clones have been identified for both CPa1 and CPa2. We are currently using a liquid culture B-Galactosidase assay for quantitative analysis of the protein interactions. The clones with the strongest interactions, correlating with the highest B-Galactosidase activity, will be sequenced and analyzed using a Bioinformatic approach.
Identification of Proteins Interacting with the Alpha Subunits of Actin Capping Protein
CSU 203
Capping protein (CP) is an actin binding protein composed of an alpha and a beta subunit, which is important for actin assembly and cell motility. Whereas lower organisms have one gene and one isoform of each subunit, higher organisms have multiple alpha isoforms (a1, a2 and a3) with conserved sequences defining highly conserved subfamilies. CPa1 and CPa2 share >90% sequence identity and the regions of divergence are highly conserved among the subunits across vertebrate. In addition, CPa1 and CPa2have distinct expression patterns in murine tissues. Endothelial cells contain only CPa2, and erythrocytes contain almost exclusively CPa1. Most tissues have both isoforms but the ratio of alpha1:alpha2 varies widely. The highly conserved sequence conservation and distinct expression patterns of CPa1 and CPa2 support our hypothesis that the CP alpha isoforms have conserved, unique and essential roles in vertebrates and will therefore, interact with unique proteins. We executed a yeast two-hybrid screen to identify proteins that interact with CPa1 and CPa2. To date, approximately 100 clones have been identified for both CPa1 and CPa2. We are currently using a liquid culture B-Galactosidase assay for quantitative analysis of the protein interactions. The clones with the strongest interactions, correlating with the highest B-Galactosidase activity, will be sequenced and analyzed using a Bioinformatic approach.
Recommended Citation
Bennett, Ryan. "Identification of Proteins Interacting with the Alpha Subunits of Actin Capping Protein." Undergraduate Research Symposium, Mankato, MN, April 2, 2019.
https://cornerstone.lib.mnsu.edu/urs/2019/oral-session-07/3
