The Biochemical Characterization of Plastin-3 (PLS3) variants linked to Congenital Diaphragmatic Hernia (CDH)

Abstract

Congenital Diaphragmatic Hernia (CDH) is a severe congenital disease characterized by an incomplete formation of the diaphragmatic septum leading to mis-localization of the abdominal organs (e.g., the liver, stomach, and bowel) to the chest cavity. Through a combination of clinical and genetic analysis, eight novel mutations in plastin-3 (PLS3), an actin-binding and bundling protein, were linked to CDH. PLS3 has two actin-binding domains, ABD1 and ABD2, which determine how the protein binds and bundles actin filaments (F-actin). The successful binding and bundling of F-actin are essential for the cell's normal functions. If the ability of PLS3 to bind and/or bundle F-actin is impaired by pathological mutations, the cell’s morphological features and critical functions will be severely altered. The CDH-linked PLS3 mutations studied in this project are localized at different domains of the protein, and their effects on the protein structure and function are characterized through high- and low-speed co-sedimentation assays, fluorescence anisotropy (FA), and differential scanning fluorimetry (DSF). All mutated plastins displayed a high degree of destabilization, when characterized through DSF, and significantly increased F-actin bundling activity, compared to wild-type (WT) protein.Thomas J Byers Memorial Scholarship, The Ohio State University Molecular Genetics DepartmentArts and Sciences Undergraduate Research Scholarship, The Ohio State University Arts and Sciences Honors CommitteeA three-year embargo was granted for this item.Academic Major: Molecular Genetic