Trying to understand the mechanisms of intracellular and extracellular cell signaling and communication, functions of multi-protein signaling complexes, protein-protein interactions and PTMs, using genetically tractable eukaryotic models, such as Slime Mould Social Ameba Dictyostelium discoideum and other conventional genetics models up to Humans. Especially interested in certain post-translational modifications of signaling proteins, such as SUMOylation and Ubiquitination. I would like to study the biological functions of these dynamic and reversible modifications, how they affect composition of these complexes, localization and activities of protein kinases, associated proteins, DNA, RNA. These functions are executed by the substrate-bound, substrate-specific enzymes, namely, E3 Ubiquitin Ligases and SUMO Ligases working in coordination with their cognate E1 and E2 enzymes. Set to apply tools of molecular genetics (genomics), biochemistry (MS/proteomics) and cell biology (high resolution fluorescent microscopy), as well as genome editing to address these questions. This work started from my EMBO postdoc at UC San Diego, and I would like to continue this work independently.
Animation:
This noncommercial video was made by HHMI Biointeractive to show how typical CRISPR-Cas9 gene editing works "to edit" specific region of genomic DNA, introducing a point mutation.
The goal: would like to apply Genome Editing (especially, Prime and Base Editing): (1) to make nucleotide substitutions in genomic DNA that encode lysines in the substrate proteins, removing previously mapped sites of SUMOylation; (2) targeting other sequence polymorphisms that are expected to have an impact on SUMOylation; (3) attempt engineering novel "synthetic" artificial SUMOylation sites, predicted to affect the activities of the target substrate proteins; (4) developing "knock-in" strategy to insert "in frame" fluorescent reporters ad/or peptide tags, so that resulting proteins could be analyzed, using specific antibodies.