Spliceosomal Processes in Nucleomorphs

ongoing

"The ""Splicing Processes in Nucleomorphs"" (SPiN) project leverages the unique properties of nucleomorphs to explore in situ the nuclear architecture and gene expression at an unprecedented level. Using cutting-edge cryogenic electron microscopy (cryo-EM) techniques such as tomography, subtomogram averaging and single particle analysis, I want to provide novel understanding of how pre-mRNA splicing happens in the context of genome reduction. I will overcome the limitations of current cryo-ET studies of the nucleus that struggle with the large size and dense environment of typical eukaryotic nuclei. Nucleomorphs, with their small size, enable a more accessible and efficient approach to in situ studies, bypassing the need for complex sample preparation steps. The SPiN project is structured around three specific aims: (1) isolating intact nucleomorphs for comprehensive proteomics and transcriptomics and structural analysis, (2) purifying and structurally characterizing spliceosome complexes unique to nucleomorphs, and (3) visualizing the structural landscape of nucleomorphs to understand the spatial organization of gene expression targeting spliceosome structures for in situ structural determination. By establishing nucleomorphs as a powerful model for in situ studies, this project promises to uncover new insights into pre-mRNA splicing mechanisms and nuclear organization, with far-reaching implications across evolutionary biology, cell biology, and biotechnology. The potential for groundbreaking discoveries positions nucleomorphs as a transformative tool to advance our understanding of eukaryotic genome evolution and the mechanisms underlying nuclear processes."