Fig. 7

MAPH-1.1 is a microtubule-associated protein related to mammalian MAP1 proteins. a Sequence similarity between MAPH-1.1 and human MAP1A, and between MAPH-1.1 and Drosophila Futsch. For each protein the predicted disordered region is indicated in green. Protein sizes are to scale. For MAP1A and Futsch, a red line indicates the proteolytic cleavage site that is used to generate a light chain (LC2 and LCf). A dotted red line indicates the homologous position in MAPH-1.1. Sequence similarity is indicated for three regions: two conserved N- and C-terminal regions (indicated in gray) and the intervening less conserved region. Amino acid coordinates for these regions are: MAP-1 N-terminal region 274–544, MAP-1 C-terminal region 2847–3041, MAPH-1.1 N-terminal region 1–230, MAPH-1.1 C-terminal region 742–878, Futsch N-terminal region 291–956, and Futsch C-terminal region 5082–5495. b Phylogenetic tree of MAP1-related proteins. Color coding indicates groups containing human MAP1A (green), MAP1B (red), and MAP1S (blue). c Western blots showing co-immunoprecipitation of DGL-1::GTA and GFP::MAPH-1.1. Input lysates (150 μg protein, lanes 1–4) show expression of DLG-1::GTA (arrowheads), GFP::MAPH-1 (arrow), and the empty GTA tag (asterisk). MAPH-1.1 co-purifies with DLG-1::GTA (lane 6) but not with the GTA tag alone (lane 5). Strains used are N2 (lane 1), BOX188 (lane 2), BOX209 (lanes 3 and 5), BOX212 (lanes 4 and 6). d Expression of GFP::MAPH-1 from an endogenously tagged GFP::maph-1.1 locus. Nerve ring, pharynx, seam cells, and hypodermis panels were taken from the same L4 stage animal. Body wall muscle and vulva panels are ventral views from a second animal. Arrowheads indicate the nerve ring an two seam cells. Cartoons are schematic representations of the areas imaged. e Expression of an N-terminal GFP::MAPH-1.1 fusion in 6-day-old primary rat hippocampal neuron cultures. Cells were co-stained for MAP2, which localizes to microtubules in dendrites, and for α-tubulin. The right panel is a merge with MAP2 in cyan, MAPH-1.1 in green, and microtubules in magenta. The arrowhead indicates the axon. f Expression of a C-terminal DLG-1::mCherry fusion in 24-day-old primary hippocampal neuron cultures. All scale bars are 10 μm