Prenylation-deficient G protein gamma subunits disrupt GPCR signaling in the zebrafish.
Prenylation of G protein gamma (gamma) subunits is necessary for the membrane localization of heterotrimeric G proteins and for functional heterotrimeric G protein coupled receptor (GPCR) signaling. To evaluate GPCR signaling pathways during development, we injected zebrafish embryos with mRNAs encoding Ggamma subunits mutated so that they can no longer be prenylated. Low-level expression of these prenylation-deficient Ggamma subunits driven either ubiquitously or specifically in the primordial germ cells (PGCs) disrupts GPCR signaling and manifests as a PGC migration defect. This disruption results in a reduction of calcium accumulation in the protrusions of migrating PGCs and a failure of PGCs to directionally migrate. When co-expressed with a prenylation-deficient Ggamma, 8 of the 17 wildtype Ggamma isoforms individually confer the ability to restore calcium accumulation and directional migration. These results suggest that while the Ggamma subunits possess the ability to interact with G Beta (beta) proteins, only a subset of wildtype Ggamma proteins are stable within PGCs and can interact with key signaling components necessary for PGC migration. This in vivo study highlights the functional redundancy of these signaling components and demonstrates that prenylation-deficient Ggamma subunits are an effective tool to investigate the roles of GPCR signaling events during vertebrate development.
Carnegie Institution, Department of Embryology, Johns Hopkins University, Baltimore, MD 21218, United States.
Timothy
T
Mulligan
Heiko
H
Blaser
Erez
E
Raz
Steven A
SA
Farber
D000818:Animals; D002118:Calcium / Q000378:metabolism; D002465:Cell Movement; D004625:Embryo, Nonmammalian / Q000378:metabolism; D047108:Embryonic Development; D044387:GTP-Binding Protein beta Subunits / Q000378:metabolism; D044388:GTP-Binding Protein gamma Subunits / Q000032:analysis / Q000235:genetics / Q000378:metabolism*; D005854:Germ Cells / Q000378:metabolism; D017368:Protein Prenylation; D012333:RNA, Messenger / Q000378:metabolism; D043562:Receptors, G-Protein-Coupled / Q000378:metabolism*; D015398:Signal Transduction; D015027:Zebrafish / Q000196:embryology* / Q000378:metabolism
D044387:GTP-Binding Protein beta Subunits; D044388:GTP-Binding Protein gamma Subunits; D012333:RNA, Messenger; D043562:Receptors, G-Protein-Coupled; D002118:Calcium