A tool to pulse-label yeast Nuclear Pore Complexes in imaging and biochemical experiments

Abstract

Nuclear Pore Complexes (NPCs) are key gateways to the nucleus and major organizers of genome architecture. Despite their importance, it is still not fully understood how NPCs are formed and degraded. Tools to track specific NPCs over time or under stress could unlock critical insights into these questions. Here, we demonstrate that a brief pulse of expression of a previously developed nanobody against bakers yeast nucleoporin Nup84 (Nordeen et al. 2020) enables a robust, rapid, and straightforward method for pulse-labeling NPCs in both imaging and affinity purification experiments. This approach offers an alternative to permanent yet less rapid genetic fluorophore- or tag-switching techniques and provides a powerful tool for studying NPC inheritance and turnover through both microscopy and biochemical methods.