An Amphipathic Helix in Brl1 is Required for Nuclear Pore Complex Biogenesis in S. cerevisiae - KARMA analysis of Brl1 (DIA)

This project was aimed at analyzing the binding of Brl1 to the nuclear pore complex during the assembly process using the Kinetic Analysis of incorporation Rates in Macromolecular Assemblies (KARMA, Onischenko et al. 2020) method. In brief, we affinity purified Brl1 at several time points following SILAC metabolic labeling in growing S. cerevisiae cultures. Then, we analyzed the metabolic labeling for all co-purified nucleoporins and used these labeling kinetics to infer the timing of Brl1 binding during the native nuclear pore complex assembly.