Ac3/Ac-Associated Protein 1 Complex Regulates Actin Filament Dynamics In Pancreatic

Abstract

Adenylyl cyclase-associated protein 1 (CAP1) is a scaffolding protein involved in the regulation of actin filament formation and cell motility. CAP1 plays a role in the motility of pancreatic cancer cells. Using co-immunoprecipitation, our data showed an upregulation of adenylyl cyclase 1 and 3, and an association with CAP1 in pancreatic cancer cell lines, HPAC and PANC-1. The goal of this project was to study whether CAP1 links AC1 and AC3 to globular actin in pancreatic cancer cells. AC1 or AC3 was knocked down using small interference RNA. After stimulation with forskolin, a direct stimulator of adenylyl cyclase, HPAC and PANC-1 cells were lysed. Total lysate was rotated with anti-actin antibody followed by complex collection with an immuno- precipitation reagent. The immunocomplexes were probed with anti-CAP1 antibody in western-blotting. The results showed that in scrambled siRNA cells stimulated with forskolin, the association between CAP1 and actin was not affected. The CAP1-actin complex was impaired in AC3-deficient cells, but was not modified in AC-1 deficient cells. In conclusion, CAP1 acts as a scaffolding protein by holding the complex between G-actin and AC3, but not AC1. Funding Source: Center for Undergraduate Research & Scholarship, Department of Biological Sciences, Scholarly Activity Award