WormBase Tree Display for Picture: WBPicture0000009849

WBPicture0000009849 Description: Figure 3. UNC-1 and UNC-9 Colocalized in Body-Wall Muscle and the Nervous System.(A) When UNC-1::HA and Myc::UNC-9 were coexpressed specifically in body-wall muscle cells under the control of Pmyo-3, colocalization was observed at body-wall muscle intercellular junctions, muscle arms, and near the ventral and dorsal nerve cords, where muscle arms from the two ventral or dorsal quadrants interdigitate. Immunoreactivity was absent in some muscle cells because of the mosaic expression of the nonintegrated transgenes. The nerve cord is indicated by arrow heads in the merged picture. The scale bar represents 50 um.(B) When Myc::UNC-9 and UNC-1::HA were coexpressed specifically in neurons under the control of Punc-47, colocalization of the two proteins was observed along the nerve cords. The scale bar represents 20 um.(C) Coexpression of UNC-1::YFPa (UNC-1 fused to YFP amino terminal) and UNC-9::YFPc (UNC-9 fused to YFP carboxyl terminal) in body-wall muscle cells under the control of Pmyo-3 reconstituted the fluorophore of YFP in vivo. Top panel: Fluorescent puncta were observed at intercellular junctions between muscle-cell bodies and between muscle arms along the nerve cord with full-length UNC-1 and UNC-9. A selected region (marked by a rectangular frame) is enlarged and shown above the image (same for the middle panel). Middle panel: Fluorescent puncta were still observed with deletion of the amino terminal of UNC-1 (amino acid residues 1-167). Lower panel: Deletion of the carboxyl terminal of UNC-1 (amino acid residues 171-289) prevented the BiFC. The bright fluorescent signal in this image was due to autofluorescence of the gut. The scale bar represents 50 um.Note: UNC-1::HA was functional because it largely rescued the behavioral phenotype of unc-1(e719) when expressed in neurons under the control of Prab-3 (data not shown). Myc::UNC-9 appeared to be a dominant-negative protein because it caused behavioral defects similar to those of unc-9(lf) when expressed in neurons of wild-type worms under the control of Prab-3 (data not shown), which could conceivably be due to coassembling of a nonfunctional Myc::UNC-9 with wild-type UNC-9 at intercellular junctions. The UNC-9 fusion protein used for BiFC assay was probably functional because UNC-9::GFP was functional. The BiFC fusion protein used for the full-length UNC-1 might also be functional because UNC-1::GFP is functional [46]. Similar to UNC-9::GFP, UNC-9::YFPc appeared to be a gain-of-function protein because animals expressing it moved more slowly than did wild-type animals. However, the locomotion defect was not as severe as that in animals expressing UNC-9::GFP (data not shown).
  Name: F3.jpg
  Depict: Expr_pattern: Expr4819
      Expr4820
    Anatomy: WBbt:0005735
      WBbt:0005813
  Acknowledgment: Template: Reprinted from <Journal_URL>, <Article_URL>, Copyright <Publication_year>, with permission from <Publisher_URL>.
    Publication_year: 2007
    Article_URL: DOI id 10.1016/j.cub.2007.06.060
    Journal_URL: CurrentBiology
    Publisher_URL: Elsevier
  Reference: WBPaper00030879