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Analysis of GLD-1 Post Translational Modification

Faculty Mentor: Sudhir Nayak

Student: John Fang

The objective of this summer’s project was to the study the regulation of GLD-1 (defective in Germ Line Development), a RNA binding protein that is important for normal germ-line development in the model organism Caenorhabditis elegans (C. elegans). In mutant strains where GLD-1 is not expressed, hermaphroditic oogenesis ceases and the germ-line tumors form. Therefore, correct expression of GLD-1 in the germ-line of C. elegans is critical and is highly regulated by various cellular mechanisms. One suspected mechanism is the phosphorylation of the GLD-1 protein for the modulation of its functionality. In normal animals, GLD-1 protein levels gradually increase from the mitotic zone through the transition zone, and reach the highest levels of expression during the pachytene phase of meiosis. Prior to oocyte development, GLD-1 levels drop abruptly. By analyzing the phosphorylation levels of GLD-1 protein in germ-line, the relationship between phosphorylation and the levels of expression of GLD-1 may be explored. Western Blotting is common method used detect changes proteins according to mass, and may be used to analyze whether or not the phosphorylation of GLD-1 is actually utilized as a regulatory mechanism. This summer was focused on optimizing the detection protocol for phosphorylated GLD-1 versus non-phosphorylated GLD-1 isoforms. I plan on continuing this research in the future with more focus on the specific stage GLD-1 is phosphorylated as well as the specific effects of phosphorylation.