Developmental fidelity is imposed by genetically separable RalGEF activities that mediate opposing signals.
PLoS Genet. 2019 May 14;15(5):e1008056. doi: 10.1371/journal.pgen.1008056. eCollection 2019 May.
Shin H1, Braendle C2, Monahan KB3, Kaplan REW3, Zand TP3,4, Mote FS1, Peters EC3, Reiner DJ1,3,4.
1. Institute of Biosciences and Technology, Texas A&M Health Science Center, Texas A&M University, Houston, TX, United States of America.
2. Université Côte d’Azur, CNRS, Inserm, IBV, Nice, France.
3. Lineberger Comprehensive Cancer Center, University of North Carolina, Chapel Hill, NC, United States of America.
4. Department of Pharmacology, University of North Carolina, Chapel Hill, NC, United States of America.
The six C. elegans vulval precursor cells (VPCs) are induced to form the 3°-3°-2°-1°-2°-3° pattern of cell fates with high fidelity. In response to EGF signal, the LET-60/Ras-LIN-45/Raf-MEK-2/MEK-MPK-1/ERK canonical MAP kinase cascade is necessary to induce 1° fate and synthesis of DSL ligands for the lateral Notch signal. In turn, LIN-12/Notch receptor is necessary to induce neighboring cells to become 2°. We previously showed that, in response to graded EGF signal, the modulatory LET-60/Ras-RGL-1/RalGEF-RAL-1/Ral signal promotes 2° fate in support of LIN-12. In this study, we identify two key differences between RGL-1 and RAL-1. First, deletion of RGL-1 confers no overt developmental defects, while previous studies showed RAL-1 to be essential for viability and fertility. From this observation, we hypothesize that the essential functions of RAL-1 are independent of upstream activation. Second, RGL-1 plays opposing and genetically separable roles in VPC fate patterning. RGL-1 promotes 2° fate via canonical GEF-dependent activation of RAL-1. Conversely, RGL-1 promotes 1° fate via a non-canonical GEF-independent activity. Our genetic epistasis experiments are consistent with RGL-1 functioning in the modulatory 1°-promoting AGE-1/PI3-Kinase-PDK-1-AKT-1 cascade. Additionally, animals lacking RGL-1 experience 15-fold higher rates of VPC patterning errors compared to the wild type. Yet VPC patterning in RGL-1 deletion mutants is not more sensitive to environmental perturbations. We propose that RGL-1 functions to orchestrate opposing 1°- and 2°-promoting modulatory cascades to decrease developmental stochasticity. We speculate that such switches are broadly conserved but mostly masked by paralog redundancy or essential functions.