title: Mutational analysis of the Caenorhabditis elegans ankyrin gene unc-44 demonstrates that the large spliceoform is critical for neural development.
abstract: In Caenorhabditis elegans, unc-44 mutations affect axonal outgrowth and guidance, leading to locomotory defects. The wild-type unc-44 gene encodes a family of ankyrin proteins, which, in addition to the conventional ankyrins, includes a novel ankyrin isoform with an extended C-terminal domain, referred to AO13 ankyrin. Six spontaneous unc-44 mutations and their reversions were analyzed in order to localize regions critical for gene function. The q331::Tc1 and rh1013::Tc1 mutations were mapped to the portion of the gene encoding the conventional ankyrins, mn339 had an uncharacterized 2-kb insertion in the serine/threonine/glutamic acid/proline-rich (STEP) repeat block 5, st200::Tc5(variant) and rh1042::Tc1 were localized near the C-terminus, and mn259 resulted from two Tc1 insertions, one in STEP block 6 and the other near the C-terminus. Tc1 excisions in several revertants resulted either in the restoration of the wild-type sequence, or were associated with small in-frame deletions or insertions. Reversion of mn339 resulted in the net excision of 2463 bp of genomic DNA, including the region encoding parts of STEP blocks 5 and 6 and the intervening hydrophobic region. Interestingly, additional Tc1 insertions at a 5' exon/intron boundary were found in revertants of st200 and rh1042. Reversion of the st200::Tc5 mutation resulted in excision of the Tc5 element, and the insertion of two copies of Tc1 at different sites. The wild-type unc-44 gene produces multiple transcripts - shorter RNAs determined to be approximately 1, 3.2, 5, 6, and 7 kb long, and two large transcripts estimated to be 22 and 26 kb in length. The largest transcripts were affected by all unc-44 mutations and are proposed to be essential for axonal outgrowth and guidance.
authors.affiliation: Department of Biological Sciences, Campus Box 4120, Illinois State University, Normal, IL 61790-4120, USA.
authors.forename: P
authors.initials: P
authors.lastname: Boontrakulpoontawee
authors.forename: A J
authors.initials: AJ
authors.lastname: Otsuka
mesh_terms: D017398:Alternative Splicing; D000818:Animals; D017487:Ankyrins / Q000235:genetics*; D001369:Axons / Q000378:metabolism; D017173:Caenorhabditis elegans / Q000196:embryology / Q000235:genetics*; D029742:Caenorhabditis elegans Proteins; D004252:DNA Mutational Analysis; D009420:Nervous System / Q000166:cytology / Q000196:embryology / Q000378:metabolism; D017422:Sequence Analysis, DNA
chemical_list: D017487:Ankyrins; D029742:Caenorhabditis elegans Proteins; C093514:unc-44 protein, C elegans