Tanja Bange (Presenting Author) | N-terminal processing and degradation | Institute of Medical Psychology, Goethstr. 31, 80336, Munich, Germany
Abstract
N-terminal acetylation (Nt-acetylation) is a ubiquitous protein modification with key roles in protein stability, folding, and complex formation. NatA, the major N-terminal acetyltransferase in human cells, modifies a large portion of the proteome, yet its global functional impact remains insufficiently characterized. To investigate NatA-dependent processes, we engineered a CRISPR/Cas9-edited DLD-1 cell line in which the NatA auxiliary subunit NAA15 is fused in-frame with an FKBP12F36V degron (V1 tag). This allows for ligand-dependent, rapid degradation of NAA15, enabling acute and reversible perturbation of NatA function. By combining this system with pulsed stable isotope labeling by amino acids in cell culture (pSILAC), we monitor dynamic changes in protein synthesis and degradation across the proteome. This integrated approach provides a powerful tool to dissect the co- and post-translational roles of NatA in proteostasis and to identify candidate substrates and pathways sensitive to N-terminal acetylation.
