Mutations in a C. elegans Gqalpha gene disrupt movement, egg laying, and viability - PubMed (original) (raw)

Mutations in a C. elegans Gqalpha gene disrupt movement, egg laying, and viability

L Brundage et al. Neuron. 1996 May.

Abstract

We find that C. elegans egl-30 encodes a heterotrimeric G protein a subunit more than 80% identical to mammalian Gqalpha family proteins, and which can function as a Gqalpha subunit in COS-7 cells. We have identified new egl-30 alleles in a selection for genes involved in the C. elegans acetylcholine response. Two egl-30 alleles specify premature termination of Gqalpha and are essentially lethal in homozygotes. Animals homozygous for six other egl-30 alleles are viable and fertile, but exhibit delayed egg laying and leave flattened tracks. Overexpression of the wild-type egl-30 gene produces the opposite behavior. Analysis of these mutants suggest that their phenotypes reflect defects in the muscle or neuromuscular junction.

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Figures

Figure 1. A C. elegans Homolog of Gqα and G11α

(A) The deduced amino acid sequence of C. elegans Gqα is shown compared with Gqα (82% identical) and G11α (83% identical) from mouse. Sequences were aligned using the program Pileup (GCG). Dashes indicates amino acid identities, and dots indicate gaps. (B) Structure of the C. elegans Gqα gene, egl-30. The exons end in the middle or end of the codon for the following amino acids: exon 1, Gly (40); exon 2, Gln (82); exon 3, Tyr (155); exon 4, Arg (198); exon 5, Glu (241); exon 6, Gly (293); exon 7, Asp (329); exon 8, Val (355).

Figure 2. C. elegans Gqα Can Activate Phospholipase C

(A) C. elegans Gqα can stimulate endogenous phospholipase C in COS-7 cells. (B) C. elegans Gqα can be activated by an a adrenergic receptor. The data shown is the average of duplicate wells, and is representative of two other similar experiments. Units are cpm of [3H]inositol phosphates released. Assay of GoAα and Gi3α were performed in a separate experiment.

Figure 3. egl-30 Hermaphrodites Are Defective in Egg Laying

Wild-type (A) and egl-30(ad805) (B) animals 28 hr after L4 larval stage. Embryos that have started to undergo morphogenesis are visible inside egl-30 but not wild-type animals. Scale bar represents 0.2 mm. egl-30 homozygotes (C) and heterozygotes (D) retain eggs. At least 30 homozygotes and 7 heterozygotes of each allele were scored. The percentage of those eggs that were late stage (1 1/2 fold stage or later) is indicated above the bars in (C). No late stage eggs were found in (D). Homozygotes of egl-30(ad810) were not tested in this assay because of lethality and sterility. syEx125 in (C) is a multicopy extra-chromosomal array overexpressing the wild-type egl-30 gene. syEx125 suppresses the egg retention of egl-30(ad809), consistent with our identification of the Gqα gene as egl-30.

Figure 4. Mutations in egl-30 Disrupt Coordinated Movement

egl-30 mutations display reduced level of movement and leave flattened sinusoidal tracks. Single wild-type (A), egl-30(ad805) (B), or dpy-20(e1282); egl-30(syEx125) (C) adult hermaphrodites were placed on 2-day-old lawns of E. coli strain OP50, and allowed to swim for 1 hr. A portion of the tracks they left are shown. Scale bar represents 0.5 mm. (D) Movement level of selected egl-30 strains. Each bar represents the average of at least seven determinations.

Figure 5. egl-30 Mutations Disrupt the Response of Egg-Laying Muscles to Serotonin and Imipramine

_egl-30_hermaphrodites lay a reduced number of eggs in serotonin (A) and imipramine (B). At least 30 unbloated animals were assayed for each drug and allele.

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References

1. Alfonso A, Grundahl K, McManus JR, Rand JB. Cloning and characterization of the choline acetyltransferase structural gene (cha-1) from C. elegans. J Neurosci. 1994;14:2290–2300. - PMC - PubMed
1. Aroian RV, Levy AD, Koga M, Ohshima Y, Kramer JM, Sternberg PW. Splicing in Caenorhabditis elegans does not require an AG at the 3′ splice acceptor site. Mol Cell Biol. 1993;13:626–637. - PMC - PubMed
1. Avery L. The genetics of feeding in Caenorhabditis elegans. Genetics. 1993a;133:897–917. - PMC - PubMed
1. Avery L. Motor-neuron M3 controls pharyngeal muscle relaxation timing in Caenorhabditis elegans. J Exp Biol. 1993b;175:283–297. - PubMed
1. Avery L, Horvitz HR. A cell that dies during wild-type C. elegans development can function as a neuron in a ced-3 mutant. Cell. 1987;51:1071–1078. - PMC - PubMed

Mutations in a C. elegans Gqalpha gene disrupt movement, egg laying, and viability - PubMed (original) (raw)