BioID-screening reveals the Alk “proximitome” of the Drosophila brain

ABSTRACT: Anaplastic lymphoma kinase (Alk) is an evolutionary conserved receptor tyrosine kinase of the insulin receptor family. In addition to its well-studied role in cancer, numerous studies on invertebrate and vertebrate models reveal that Alk-signaling is associated with a variety of complex traits such as: regulation of growth and metabolism, hibernation, regulation of neurotransmitters, synaptic coupling, axon targeting, decision making, memory formation and learning, alcohol use disorder, as well as steroid hormone metabolism. To shed light on ALK-driven signaling processes, we used BioID-based in vivo proximity labeling to reveal the molecules that interact with Alk in the Drosophila CNS. Therefore, we generated first and next generation BioID fusion proteins in the endogenous Alk locus by CRISPR/Cas9 induced homology-directed repair (HDR) resulting in viable and fertile flies. Our results show (i) that the next generation of BioID proteins (TurboID and miniTurbo) outperform the first generation BirA* fusion in terms of labeling efficiency, and (ii) allow identification of proximitomes without overexpression which are likely to be more physiologically relevant. LC-MS/MS-based BioID screening of the larval brain revealed an extensive neuronal Alk-proximitome identifying numerous novel potential components of Alk signaling complexes. Validation of Alk-proximitome candidates further revealed co-expression with Alk in the CNS.

INSTRUMENT(S):

ORGANISM(S): Drosophila Melanogaster (fruit Fly)

TISSUE(S): Brain

SUBMITTER: Proteomics Core Facility  

LAB HEAD: Ruth Palmer

PROVIDER: PXD026196 | Pride | 2025-08-22

REPOSITORIES: Pride