Logan Fickes | 2025 I.S. Symposium

Name: Logan Fickes
Title: Investigating Nab2’s Interactions with the Spliceosomal Protein SmB
Major: Neurobiology
Advisors: Seth Kelly; James West

RNA splicing is a critical cellular process carried out by the spliceosome, a complex of small nuclear ribonucleoproteins (snRNPs) and associated proteins. Disruptions in splicing are linked to many diseases, including spinal muscular atrophy, cancer, and intellectual disabilities. Nab2, and RNA-binding protein (RBP) orthologous to human ZC3H14, has been implicated in splicing regulation, but its precise role remains unclear (Soucek et al., 2016). Since ZC3H14 physically interacts with spliceosomal components, it was hypothesized that Nab2 physically interacts with the spliceosome (Soucek et al., 2016). To address this, co-immunoprecipitation (co-IP) assays were employed to test for physical interactions between Nab2-FLAG and SmB, a core spliceosomal protein, in Drosophila (Huntriss et al., 1993). The results demonstrated that Nab2-FLAG was successfully immunoprecipitated, confirming its presence in the lysate and elution fractions, but SmB was not detected in the elution lanes with Nab2-FLAG, indicating no interaction between these two proteins. These findings suggest that the co-IP methodology needs further optimization or that Nab2’s influence on splicing is likely mediated through indirect downstream effects of its other known functions. This challenges the assumption of functional overlap between Nab2 and ZC3H14 and highlights the need to reassess Nab2’s role in splicing regulation. Broader implications include a better understanding of how RBPs like Nab2 contribute to mRNA processing, with potential applications in modeling intellectual disabilities linked to ZC3H14 dysfunction in Drosophila. Future research should explore the downstream effects of Nab2’s functioning and how these might affect splicing regulation.

Posted in Symposium 2025 on May 1, 2025.