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. 2006 Nov 30;25(24):5884–5895. doi: 10.1038/sj.emboj.7601458

Figure 3.

Figure 3

GPA-12 (Q205L) stimulation of ACh release requires RHGF-1, DAG, and UNC-13. (A, B) The RGS RhoGEF mutation (rhgf-1(ok880)) completely suppressed aldicarb hypersensitivity caused by the hs∷GPA-12 (Q205L) transgene (hs∷GPA-12*;rhgf-1(ok880)) in the absence of heat shock (A) and strongly suppresses in the presence of heat shock (B). In hs∷GPA-12 (Q205L); rhgf-1(ok880) animals, expression of RHGF-1 from the cholinergic motor neuron-specific acr-2 promoter (N∷RHGF-1) at low copy number (injected at 10 ng/μl, hs∷GPA-12*;rhgf-1(ok880);N∷RHGF-1) had no effect on the response of animals to aldicarb in the absence of heat shock (A), but restored aldicarb hypersensitivity caused by heat-shock-induced expression of GPA-12 (Q205L) (B). (C) A transgene with a 10-fold increase in p.acr-2∷RHGF-1 (injected at 100 ng/μl N∷RHGF-1 × 10) caused aldicarb hypersensitivity in hs∷GPA-12*;rhgf-1(ok880) animals without heat shock (−hs). Aldicarb hypersensitivity was decreased slightly upon heat-shock-induced expression of GPA-12 (Q205L) (+hs) and much more strongly by removal of all GPA-12 using the gpa-12(pk322) mutation. (D) Inhibition of endogenous RHO-1 by heat shock expression of C3 transferase (hs∷C3T +hs) blocked aldicarb hypersensitivity caused by N∷RHGF-1 × 10 (compare rhgf-1(ok880);N∷RHGF-1 × 10;hsC3T in the absence (−hs) or presence (+hs) of C3 transferase expression). (E, F) Replacement of endogenous UNC-13 by a mutant UNC-13S unable to bind diacylglycerol (DAG) (unc-13(s69); UNC-13S (H173K)) caused aldicarb resistance (E) that is not altered by heat shock expression of constitutively active GPA-12 (Q205L) (F).