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Comments on Bastiani CA et al. (1997) International C. elegans Meeting "Genetic analysis of Gq alpha mediated signaling pathways in C. elegans" (0)
Overview
Bastiani CA, Brundage LA, Sternberg PW, & Simon MI (1997). Genetic analysis of Gq alpha mediated signaling pathways in C. elegans presented in International C. elegans Meeting. Unpublished information; cite only with author permission.
egl-30 encodes a protein with more than 82% amino acid sequence identity to mammalian heterotrimeric G protein alpha subunits of the Gq family. The mammalian Gq family can be activated by at least thirty different receptors, including receptors which respond to the neurotransmitters acetylcholine, serotonin, and glutamate. The Gq family represents a major component of G protein mediated signaling through phosphoinositides by direct activation of phospholipase C beta isoforms. Previously characterized reduction-of-function mutations have implicated egl-30 in the regulation of behaviors such as egg-laying, coordinated movement, and viability in C. elegans.1 We have engineered a constitutively active allele of egl-30 fused to the hsp-16-2 promoter element and integrated this gene fusion into the C. elegans genome. Within two hours after a heatshock regimen at 32oC for thirty minutes, these worms hypercontract and become paralyzed. Interestingly, this effect closely mimics the initial response of wild-type worms to arecoline, an acetylcholine agonist. To define downstream components of egl-30 mediated signaling pathways in C. elegans, we are screening for mutations which can suppress the paralysis which results from overexpression of the constitutively active allele of egl-30. In a screen of ~9,500 haploid genomes, we have identified 7 mutations that suppress both hypercontraction and paralysis. Four of the suppressor lines are phenotypically wild-type in the absence of heat-shock. Three of the suppressor lines are moderately to severely Dpy and become progressively bloated and sluggish as they age. We are currently in the process of carrying out additional mutagenesis screens and of mapping and further characterizing these suppressor genes. 1 L. Brundage, L. Avery, A. Katz, U.-J. Kim, J.E. Mendel, P.W. Sternberg, M.I. Simon, Neuron 16, 999-1009 (1996).
Affiliation:
- Division of Biology 147-75 California Institute of Technology Pasadena, CA 91125 U.S.A.