New Semester
Since cells harboring the mutant version of Mex67 (export factor) and lacking Upf1 (NMD factor) demonstrate high molecular weight aggregates, we are directed towards determining if Mex67p can exist in a prion like form. In order to further characterize this behavior we will verify the Q/N content of the protein and look for phenotypes that can be reversed in the presence of Guanidine HCl (GuHCl) which will indicate possible prion disruption.
Experimental Design:
Using yeast genetics, we will verify phenotypes that can be used as markers for prion disruption in the presence of GuHCl. These phenotypes may include:
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- temperature sensitivity
- drug sensitivity (cycloheximide or paromomicin)
- colony size
Additional assays for scoring the hability of mex67-5 to form prions is the cytoduction experiment. These experiments will determine if the molecular weight aggregates have a non mendelian inheritance pattern tipical of prion proteins.
