Alexander Kehl
Year: 2018
Faculty Advisor: Stephane Bentolila

“Determining the Localization of the OZ Protein Family”


Project Summary:

The goal of the experiment is to determine the subcellular localization of the proteins encoded by the genes OZ2, OZ3 and OZ4. RNA editing in vascular plants converts cytidine into uridine by a deaminase reaction. The function of RNA editing is to correct mutations in the genomes of the chloroplast and mitochondria as a part of post-transcriptional processing.  When the OZ1 gene is mutated by knocking out the OZ1 amino acid sequence, the efficacy of editing various chloroplast transcripts was affected. The plastids were transfected into Arabidopsis protoplasts and observed using fluorescence microscopy. Additionally, the plasmid was transformed into Agrobacterium and agroinfiltration was performed on the leaf of Nicotiana benthamiana. It is important to understand the localization of the OZ porteins to determine if the other members of the OZ family are affecting RNA editing in the chloroplast or in the mitochondria. In addition, the experiment supports the idea of the proteins being localized at the nucleoid of the organelle. The results from the experiment provides a foundation for future knock-out experimentation on the OZ proteins. Knock-out experimentation is essential to determine the effect of the OZ proteins on the RNA editosome. There was found to be a variance in the distribution of localization between the different proteins in the OZ family. The location of the proteins was convergent on points of interaction between the OZ protein and an unknown target molecule that is thought to be the nucleoid of the organelle.

My Experience:

Over the summer I learned and expanded on a wide variety of techniques in molecular and plant biology. A large portion of the lab work was done independently, along with work assisted, and explained by my mentor. In my personal experience I received guidance in professional development to better prepare me for my scientific career. I was also exposed to a wide range of research topics in the field of plant biology and related disciplines.