To study yeast actin cytoskeleton involved in endocytosis and actin cable using optigenetics
Jebarani, Jebamony Jasmin
Date of Issue2016-05-25
School of Chemical and Biomedical Engineering
Actins in eukaryotic organisms plays a major role in cellular functions. They primarily appear in two major forms i.e., actin patch and actin cable. Actin patch plays an important role in endocytosis by providing the force that is needed for the plasma membrane invagination and engulfment of the cargo. Actin patches are nucleated by Arp2/3 complex which in turn is activated by Las17. On the other hand, actin cable plays vital role in transporting the cargo into and within the cell with the help of motor protein called Myo2. Actin cables are nucleated by formins called Bni1 and Bnr1. Optogenetics is a powerful tool in order to study the dynamics of a protein. It can control the expression of a gene at genetic level. Two light-sensitive proteins PhyB and PIF, when fused with fluorescent proteins and in the presence of PCB at 650nm, they form complex and the process can be reversed by shining 750nm. Here, I studied the role of four proteins viz., Las17, Bni1, Myo2 and Cmd1 in endocytosis and actin cable polymerization. Las17 and Bni1 plays a major role in endocytosis, actin cable polymerization and their inhibition affects the rate of endocytosis and the velocity of actin cable polymerization significantly. Myo2 and Cmd1 genes also play a significant role in endocytosis and actin cable polymerization.