Differential requirement for the nonhelical tailpiece and the C terminus of the myosin rod in Caenorhabditis elegans muscle.
Myosin heavy chain (MHC) is a large, multidomain protein important for both cellular structure and contraction. To examine the functional role of two C-terminal domains, the end of the coiled-coil rod and the nonhelical tailpiece, we have generated constructs in which residues within these domains are removed or mutated, and examined their behavior in Caenorhabditis elegans striated muscle. Genetic tests demonstrate that MHC lacking only tailpiece residues is competent to support the timely onset of embryonic contractions, and therefore viability, in animals lacking full-length MHC. Antibody staining experiments show that this truncated molecule localizes as wild type in early stages of development, but may be defective in processes important for thick filament organization later in embryogenesis. Ultrastructural analysis reveals thick filaments of normal morphology in disorganized arrangement, as well as occasional abnormal assemblages. In contrast, molecules in which the four terminal residues of the coiled coil are absent or mutated fail to rescue animals lacking endogenous MHC. Loss of these four residues is associated with delayed protein localization and delayed contractile function during early embryogenesis. Our results suggest that these two MHC domains, the rod and the tailpiece, are required for distinct steps during muscle development.
Hoppe
Pamela E
PE
Washington University School of Medicine, St Louis, Missouri 63110, USA. phoppe@genetics.wustl.edu
Andrews
Rebecca C
RC
Parikh
Payal D
PD
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D017173:Caenorhabditis elegans / Q000196:embryology / Q000235:genetics* / Q000378:metabolism* / Q000648:ultrastructure; D017124:Conserved Sequence; D018509:DNA, Helminth / Q000235:genetics; D008969:Molecular Sequence Data; D009068:Movement; D018482:Muscle, Skeletal / Q000196:embryology / Q000378:metabolism* / Q000648:ultrastructure; D018995:Myosin Heavy Chains / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D015879:Myosin Subfragments / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D010641:Phenotype; D017384:Sequence Deletion
D018509:DNA, Helminth; D015879:Myosin Subfragments; D018995:Myosin Heavy Chains