A Kinome RNAi Screen Reveals Genes Required for Muscle Tissue Maintenance

Abstract

NUAK is an AMPK-related kinase which controls autophagy (cellular recycling) of an important linker protein in muscle called Filamin1. The absence of NUAK prevents the turnover of Filamin, presents an accumulation of autophagic markers such as p62 and ATG8 and leads to muscle degeneration. The goal of our research was to determine which other kinase genes might be required to regulate autophagy by turning them off with RNA interference (RNAi) and looking for abnormal muscle structure and attachment. 25 candidate kinase genes were selected and turned off in the muscle of Drosophila melanogaster larvae using RNAi. The larvae were then dissected and imaged under a laser confocal microscope to observe muscle structure. Knockdown of seven kinase genes yata, nippedA, CDK9, for, RIOK1, lic, and Vps15 showed abnormal muscle structure such a detachment and muscle thinning. These kinase genes will be further analyzed for their requirement in the proper autophagy of Filamin.