Investigation into the Akirin Subcellular Localization and Developmental Interactions

Akirin is a highly conserved nuclear protein ubiquitously expressed in nonfungal eukaryotic species and is known to be involved in many biological processes such as embryonic muscle patterning and innate immune function. Akirin functionality is essential for the prevention of disease and developmental defects. This investigation focused on both the subcellular localization of Akirin based on a putative nuclear localization signal and the genetic interaction between Akirin and the Nucleosome Remodeling Deacetylase (NuRD) complex. Through transfection of Drosophila S2 cells with Akirin mutant constructs we were able to visualize the localization patterns of Akirin and how the patterns relate to the Nuclear Localization Signal (NLS). Data shows that basic amino acids in the N-terminal region of the protein are sufficient for driving nuclear localization of the protein, though the protein isn’t localized as effectively. In a parallel experiment, we investigated Akirin’s genetic interactions and the consequential developmental consequences. Through Drosophila embryo immunostaining for Myosin Heavy Chain, Green Fluorescent Protein, and Tinman it was seen that there is an interaction between Akirin and multiple subunits of the NuRD complex as it pertains to skeletal muscle development and cardio myogenesis. The findings of this research show implications for understanding the nuclear localization patterns of the protein and the possibility of using the endogenous bipartite Akirin NLS as a cargo protein delivery system. The skeletal and cardiac phenotypes observed provide evidence for an interaction between Akirin and the NuRD complex shedding light on multiple different areas of developmental biology