Figure 2.
nsy-1 Is Expressed in the AWC Neurons and May Act Downstream of
unc-43/CaMKII. (A and B) A
nsy-1::GFP transgene with 10 kb of
nsy-1 upstream sequence was expressed in many cells, including AWC. (A) A confocal projection of an L1 larval stage animal with wide expression of
nsy-1::GFP. (B) Merged
nsy-1::GFP and Nomarski images of an L1 animal. An arrow indicates AWC.(C-F) Animals bearing an integrated
str-2::GFP reporter. Fluorescence posterior to the head is gut autofluorescence or gut expression of the transgene marker
elt-2::GFP. (C) Wild-type animals expressed
str-2::GFP in one AWC neuron. (D) Animals expressing
odr-3::NSY-1(GF) did not express
str-2::GFP in either AWC neuron. (E) An
unc-43 loss-of-function mutation caused
str-2::GFP expression in both AWC neurons. (F) An
unc-43 loss-of-function mutant expressing
odr-3::NSY-1(GF) did not express
str-2::GFP in either AWC neuron. The dorsally-located, dim-staining cells in (D) and (F) are the ASI neurons, where
str-2::GFP is sometimes weakly expressed. Dorsal is up and anterior is to the left in all panels.(G) Association between UNC-43 and NSY-1 in mammalian cells. Human embryonic 293 cells were transfected with a control vector (-); an HA-tagged, calcium-independent, activated form of UNC-43, HA-UNC-43(T284D); and a catalytically inactive, T7-tagged form of NSY-1, T7-NSY-1(K703M). Cell lysates were immunoprecipitated (IP) with anti-T7 antisera. Immunoprecipitates were immunoblotted (IB) with anti-HA (top panel) and anti-T7 (middle panel); UNC-43 was present in the NSY-1 immunoprecipitate (top panel). Whole-cell extracts were also immunoblotted with anti-HA (bottom panel).