Myosin and paramyosin of Caenorhabditis elegans embryos assemble into nascent structures distinct from thick filaments and multi-filament assemblages.
The organization of myosin heavy chains (mhc) A and B and paramyosin (pm) which are the major proteins of thick filaments in adult wild-type Caenorhabditis elegans were studied during embryonic development. As a probe of myosin-paramyosin interaction, the unc-15 mutation e73 which produces a glu342lys charge change in pm and leads to the formation of large paracrystalline multi-filament assemblages was compared to wild type. These three proteins colocalized in wild-type embryos from 300 to 550 min of development after first cleavage at 20 degrees C on the basis of immunofluorescence microscopy using specific monoclonal antibodies. Linear structures which were diversely oriented around the muscle cell peripheries appeared at 360 min and became progressively more aligned parallel to the embryonic long axis until distinct myofibrils were formed at 550 min. In the mutant, mhc A and pm were colocalized in the linear structures, but became progressively separated until they showed no spatial overlap at the myofibril stage. These results indicate that the linear structures represent nascent assemblies containing myosin and pm in which the proteins interact differently than in wild-type thick filaments of myofibrils. In e73, these nascent structures were distinct from the multi-filament assemblages. The overlapping of actin and mhc A in the nascent linear structures suggests their possible structural and functional relationship to the "stress fiber-like structures" of cultured vertebrate muscle cells.
Epstein
H F
HF
Department of Neurology, Baylor College of Medicine, Houston, Texas 77030.
Casey
D L
DL
Ortiz
I
I
D000199:Actins / Q000378:metabolism; D000818:Animals; D000911:Antibodies, Monoclonal; D017173:Caenorhabditis elegans / Q000196:embryology / Q000648:ultrastructure; D005455:Fluorescent Antibody Technique; D009132:Muscles / Q000196:embryology* / Q000648:ultrastructure; D009218:Myosins / Q000378:metabolism*; D014335:Tropomyosin / Q000378:metabolism*
D000199:Actins; D000911:Antibodies, Monoclonal; D014335:Tropomyosin; D009218:Myosins