The tropomyosin gene
tmy-1 spans approximately 13 kilo bases and includes 14 exons which encodes three tissue specific isoforms. The 5' end promoter common to CeTMI and CeTMII expresses in the body wall muscles, vulva, anus muscles and male tail muscles. Two of them having 284 amino acid residues are only different in the last 27 amino acid residues. By the sequence comparison of the last 27 amino acid residues to that of other animals, we assume that CeTMII could be the cytoplasmic type. The third isoform ceTMIII encoding 256 amino acids is expressed in the pharyngeal muscles by the promoter in the third intron. mRNA of CeTMIII was transspliced with SL1. The
tmy-1 gene is locates on the C. elegans genomic YAC grid near the right end of chromosome I, in the region of the
lev-11 gene (Kagawa et al, 1995). Levamisol is a potent agonist of acetylcholine in the worm and
lev-11 is isolated as twitchers (Lewis et al, 1980). The mutant embryos of
lev-11(
st557) having Pat phenotype show greatly reduced or undetectable staining with purified anti-tropomyosin antibody (Williams and Waterston, 1994). We determined the sequences of the exon parts of
tmy-1 gene from
lev-11 mutations. The
lev-11(
st557) mutation occurred at the splice donor site of exon 1 (GAAG GTTT at 2373 to GAAG ATTT) and results in translation termination at 2412 after adding twelve amino acid residues. Nucleotide sequence numbers follow that of the database of the
tmy-1 gene D38539. Although small sized protein from herozygous (+/st557) animal was not detected by Western analysis, our result could be the reason of Pat (paralyzed, arrested elongation at two fold) phenotype of the mutation. The
lev-11(X12) mutation occurred in exon 7 (AAG GAG at 10262 to AAG AAG) and results in amino acid substitution at 234 from Glu to Lys. This substitution give a charge change from - to + at this point. As this region is common in three isoforms, there may be functional importance of this region for calcium signaling from troponins to actins through molecular interactions. We can not detect mutation in the
lev-11(X1) and
lev-11 (
st536) at the moment. We have determined the mutation site of levamisol sensitive but anomalous unc mutation
unc-68 in the ryanodine receptor gene
ryr-1 (Sakube et al, unpublished observation). These results allows us to understand not only calcium signaling from post synaptic membrane to muscle filaments but also developmental importance of muscle formation in the worm. Kagawa et al, (1995) J. Mol. Biol. 251, 606-613. Lewis et al, (1980) Genetics 95, 905-925. Williams and Waterston, (1994) J. Ce.. Biol. 124, 457-490.