Protein-Protein Interactions of the β1 and β2 Forms of Actin Capping Protein
Location
CSU Ballroom
Start Date
21-4-2008 1:00 PM
End Date
21-4-2008 3:00 PM
Student's Major
Biological Sciences
Student's College
Science, Engineering and Technology
Mentor's Name
Marilyn C. Hart
Mentor's Department
Biological Sciences
Mentor's College
Science, Engineering and Technology
Description
Actin is a component of eukaryotic cells that plays a role in cell structure and motility. Actin Capping Protein (CP) is a protein associated with the regulation of actin microfilament dynamics and stability. CP is composed of two proteins, an alpha (α) and a beta (β) subunit. In vertebrate organisms, three β (βl, β2, β3) forms have been identified. The β1 form is the predominant form in muscle, where as the β2 is the predominant form in non-muscle; β3 functions in germ cells. Previous studies of genetically altered mice have shown that βl and β2 have different functions in mouse myocardium. We hypothesize that the different functions of βl and β2 is due to their ability to interact with different proteins. Therefore, we attempted to identify proteins that interact with each form using a yeast two hybrid genetic screen. We generated the necessary βl, β2 and α1 constructs and confirmed their orientation and sequence identity. The constructs were chemically transformed into Saccharomyces cerevisiae strain AH109 and their presence confirmed through plating on selective dropout media. Protein expression was verified via Western Blot analysis. Small scale screening between α1 and β1 or β2 of the transformed yeast has verified the feasibility of the screen. A large scale screen using a murine heart cDNA library is ongoing.
Protein-Protein Interactions of the β1 and β2 Forms of Actin Capping Protein
CSU Ballroom
Actin is a component of eukaryotic cells that plays a role in cell structure and motility. Actin Capping Protein (CP) is a protein associated with the regulation of actin microfilament dynamics and stability. CP is composed of two proteins, an alpha (α) and a beta (β) subunit. In vertebrate organisms, three β (βl, β2, β3) forms have been identified. The β1 form is the predominant form in muscle, where as the β2 is the predominant form in non-muscle; β3 functions in germ cells. Previous studies of genetically altered mice have shown that βl and β2 have different functions in mouse myocardium. We hypothesize that the different functions of βl and β2 is due to their ability to interact with different proteins. Therefore, we attempted to identify proteins that interact with each form using a yeast two hybrid genetic screen. We generated the necessary βl, β2 and α1 constructs and confirmed their orientation and sequence identity. The constructs were chemically transformed into Saccharomyces cerevisiae strain AH109 and their presence confirmed through plating on selective dropout media. Protein expression was verified via Western Blot analysis. Small scale screening between α1 and β1 or β2 of the transformed yeast has verified the feasibility of the screen. A large scale screen using a murine heart cDNA library is ongoing.
Recommended Citation
Strehler, Kevin Y. E.. "Protein-Protein Interactions of the β1 and β2 Forms of Actin Capping Protein." Undergraduate Research Symposium, Mankato, MN, April 21, 2008.
https://cornerstone.lib.mnsu.edu/urs/2008/poster-session-B/1
