Hedgehog signaling and Tre1 regulate actin dynamics through PI(4,5)P2 to direct migration of Drosophila embryonic germ cells

• Published in: Cell reports (Cambridge) 2021-03, Vol.34 (9), p.108799-108799, Article 108799
• Main Authors: Kim, Ji Hoon, Hanlon, Caitlin D., Vohra, Sunaina, Devreotes, Peter N., Andrew, Deborah J.
• Format: Article
• Language: English
• Subjects: cell migration; germ cells; GPCR; Hedgehog; PI(4,5)P2; PIP5K; Smoothened; Tre1; WASP; WIP
• ISSN: 2211-1247; 2211-1247
• DOI: 10.1016/j.celrep.2021.108799

The Tre1 G-protein coupled receptor (GPCR) was discovered to be required for Drosophila germ cell (GC) coalescence almost two decades ago, yet the molecular events both upstream and downstream of Tre1 activation remain poorly understood. To gain insight into these events, we describe a bona fide null allele and both untagged and tagged versions of Tre1. We find that the primary defect with complete Tre1 loss is the failure of GCs to properly navigate, with GC mis-migration occurring from early stages. We find that Tre1 localizes with F-actin at the migration front, along with PI(4,5)P2; dPIP5K, an enzyme that generates PI(4,5)P2; and dWIP, a protein that binds activated Wiskott-Aldrich syndrome protein (WASP), which stimulates F-actin polymerization. We show that Tre1 is required for polarized accumulation of F-actin, PI(4,5)P2, and dPIP5K. Smoothened also localizes with F-actin at the migration front, and Hh, through Smo, increases levels of Tre1 at the plasma membrane and Tre1’s association with dPIP5K. [Display omitted] •Tre1 localizes with dPIP5K, PI(4,5)P2, and F-actin and is required for their localization•Hh and its receptor Ptc accumulate in intracellular structures within migrating GCs•Smoothened also colocalizes with F-actin in GCs and is required for GC navigation•Hh, through Smo, increases Tre1 at the plasma membrane and its association with dPIP5K Kim et al. uncover molecular and cellular events upstream and downstream of the Tre1 G-protein coupled receptor (GPCR), which is required for germ cell navigation in Drosophila. Hedgehog signaling through Smoothened localizes Tre1 to activate F-actin assembly through dPIP5K, PI(4,5)P2, and WASP.