Novel insights into DAPK autophagic signalling using peptide aptamer combinatorial protein-interaction screens.

摘要

DAPK represents a relatively unique enzyme in the protein kinase superfamily whose major biological functions are linked to both autophagy and signal-mediated apoptosis. However, genetic studies have not yet uncovered how DAPK integrates into the core autophagy-related (Atg) machinery since DAPK is not present in a genetically tractable eukaryotic cell such as yeast. Furthermore, there have been no definitive DAPK binding proteins identified in metazoan systems that play a direct role in cooperating with DAPK in autophagy. We have utilized a growing concept in systems biology that invokes linear peptide-motifs as a fundamental mechanism driving protein-protein interactions and as a key switch underlying the dynamics of a signal transduction pathway. By using peptide combinatorial libraries as an assay that reflects the diversity of the linear peptide motif repertoire in the mammalian proteome, we identified microtubule-associated protein 1B (MAP1B) as a novel DAPK interacting protein that stimulates DAPK-dependent membrane blebbing and autophagy. MAP1B has previously been shown to form a functional interaction with the autophagosomal protein Atg8 (LC3). Together these studies define a genetic interaction between DAPK-MAP1B in the regulation of autophagy that may have particular relevance to cellular signalling pathways that regulate cell survival or cell death under distinct environmental stresses.