Functional characterization of LOC115098:A novel gene conserved in eukaryotic genomes
Tazebay, Uygar H.
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Loc115098 is a newly identified gene in our laboratory. The gene was firstly discovered in an unrelated study in which the cis-acting regulatory elements of a neighboring gene NIS (Sodium-Iodide Symporter) were investigated. It is composed of 4 exons. According to the computational predictions, it has rather interesting features such as having a composition of mostly charged amino acids, carrying a nuclear localization signal and being well conserved throughout the eukaryotic kingdom. Our aim in this study was to understand the function of this new gene. Firstly we confirmed that it is expressed in several different tissues, later we conducted a subcellular localization study whose results led us to hypothesize that the function of loc115098 could be related to cell cycle regulation. According to our hypothesis we decided to utilize a lower eukaryote carrying a human homologue of loc115098 which we selected to be Aspergillus nidulans. It was a very useful model organism for our study not only because it carries a very similar homologue of loc115098 but also it is an easily handled organism which is widely used for genetic studies. Firstly we compared the expression profile of loc115098 in several cell cycle mutants of A. nidulans as well as a wild type strain, the results indicated that the expression of loc115098 decreases in the restrictive temperature of cell cycle mutant strains. We also conducted subcellular localization studies in A. nidulans and the results indicated that it was mostly cytoplasmic and sometimes perinuclear. Our other aim was to utilize A. nidulans as a model for our knock-out experiments. We designed a knock-out system by the help of a sophisticated method called DoubleJoint PCR. Our first attempts supported that the knock-out of loc115098 was likely to cause lethality in the organism and therefore we improved our system to conditionally knock out the gene. Our preliminary results indicated that the knock out of loc115098 causes cell death and we suspect that it could be due to a damage in one of the crucial metabolic events for cell viability such as mitosis. Our attempts to functionally characterize the function of loc115098 are currently going on.