Project Description
Multiple abiotic and biotic factors, including a conserved dual kinase/RNase Ire1, govern the protein homeostasis (proteostasis) from yeast Saccharomyces cerevisiae to human cells. We recently showed that the Ire1-mediated proteostasis yeast involves two distinct phases: (I) an early phase and (II) a later phase. The early phase predominately operates through the Ire1. During the later phase, the Ire1 pathway is regulated by protein kinases Pkh1 and its downstream substrate kinase Ypk1. In the kinase Ypk1, Tyr-536 to Cys mutation(Y536C) is known to cause a temperature sensitive (ts)-phenotype. This residue Y536 resides at the potential substrate/effector binding site of the Ypk1 kinase domain. To identify potential effector(s), we conducted a dosage suppressor screen, which identified that over-expression of both the GTPase Rho2 and the RNase Ire1 allowed the ypk1ts strain to grow at 37oC. These results suggest that both Ire1 and Rho2 functions to activate the kinase Ypk1. Further analysis revealed that the ire1 strain exhibited strong sensitivity, whereas the rho2 strain showed moderate sensitivity to tunicamycin, which interferes with the protein folding processes in the endoplasmic reticulum. The student will work on to understand the deeper genetic link between the RNase Ire1 and kinase Ypk1 and the GTPase Rho2.
Tasks and Responsibilites
-Searching literatures
-Analyzing the sequence and structures of Ire1, Ypk1 and Rho2
-Attending the Dey Lab meeting as and when required
-Conducting molecular and biochemical studies in the Dey Lab.
Desired Qualifications
None Listed.