Journal: The Journal of Cell Biology

Article Title: The kinetically dominant assembly pathway for centrosomal asters in Caenorhabditis elegans is γ-tubulin dependent

doi: 10.1083/jcb.200202047

Figure Lengend Snippet: tbg-1(t1465) mutants have a mutation in a residue highly conserved among tubulins. (A) tbg-1(t1465) is a point mutation that leads to the substitution of alanine 401 with valine. A sequence alignment of the COOH termini of α-, β-, and γ-tubulins from S. cerevisiae and C. elegans, α- and β-tubulin from pig, and human γ-tubulin from human is shown. Amino acids conserved in 80–100% of the sequences are highlighted in yellow and alanine 401 is in red. Arrows and tubes above the alignment indicate conserved β sheets and α helices, respectively . (B–D) Characterization of the MT cytoskeleton in tbg-1(t1465) embryos. (B) An interphase tbg-1(t1465) embryo stained for MTs (left) and DNA and γ-tubulin (blue and red, right). A remnant of the spindle from the second meiotic division of the oocyte pronucleus is visible (black arrow), but no centrosomal asters are detected. γ-Tubulin localizes to centrosomes (arrows) that fail to associate with the sperm pronucleus (arrowhead). (C) Mitotic tbg-1(t1465) embryos were stained for MTs (left), γ-tubulin (middle), and DNA (right). Two typical examples are shown here (top and bottom). (D) Mitotic tbg-1(t1465 + RNAi) embryos stained for MTs and ZYG-9 (green and red, left), γ-tubulin (middle), and DNA (right). Bar, 10 μm.

Article Snippet: sas-3(t1465) mutant worms were obtained from Ralf and Heinke Schnabel (Max Planck Institut fuer Biochemie, Martinsried, Germany).

Techniques: Mutagenesis, Sequencing, Staining