Figure 1.
oig-8 Expression in Sensory Neuronsand Localization to Ciliated Endings(A)
oig-8 locus,
oig-8 promoter fusion to assessexpression,
oig-8 fosmid-based translational reportersto assess protein localization (the N-terminal fusion hasGFP inserted immediately after the signal peptidesequence; the C-terminal fusion has GFP insertedimmediately before the stop codon), CRISPR deletionallele
ot818, and nonsense mutation (Q116Amber)allele
gk867223. The OIG-8 protein contains a signalpeptide sequence (SS) and transmembrane (TM)domain.(B and C) GFP::OIG-8 (from the
oig-8fosmid::Ngfptranslational reporter, otEx6368) localizes to amphidcilia (B) and to axonal and dendritic membranes of PVD(C). Fusing GFP to the C-terminal end of the
oig-8 locus(
oig-8fosmid::Cgfp, otEx6815) results in severe sortingdefects; the OIG-8::GFP protein is stuck in the cellbody, suggesting that C-terminal residues may becritical for OIG-8 trafficking (B).(D) GFP::OIG-8 expressed in individual amphidwing neurons localizes to cilia (GFP signals are colorinverted and then transformed to black/white signals).GFP::OIG-8 from
odr-10prom::gfp::
oig-8 (otEx6936) localizes to the cilia of AWA in L4 animals (middle).GFP::OIG-8 from
str-1prom::gfp::
oig-8 (otEx6807) localizes to the cilia of AWB in L4 animals (middle).GFP::OIG-8 from ceh-36prom3::gfp::
oig-8 (otEx6922)localizes to the cilia of AWC L4 animals (bottom).GFP::OIG-8 also localizes to the cilia of amphid wingneurons in L1-stage animals (Figure S2F). The ciliaschematic is adapted from [2, 23].