The carboxyl-terminal isoforms of smooth muscle myosin heavy chain determine thick filament assembly properties.
The alternatively spliced SM1 and SM2 smooth muscle myosin heavy chains differ at their respective carboxyl termini by 43 versus 9 unique amino acids. To determine whether these tailpieces affect filament assembly, SM1 and SM2 myosins, the rod region of these myosin isoforms, and a rod with no tailpiece (tailless), were expressed in Sf 9 cells. Paracrystals formed from SM1 and SM2 rod fragments showed different modes of molecular packing, indicating that the tailpieces can influence filament structure. The SM2 rod was less able to assemble into stable filaments than either SM1 or the tailless rods. Expressed full-length SM1 and SM2 myosins showed solubility differences comparable to the rods, establishing the validity of the latter as a model for filament assembly. Formation of homodimers of SM1 and SM2 rods was favored over the heterodimer in cells coinfected with both viruses, compared with mixtures of the two heavy chains renatured in vitro. These results demonstrate for the first time that the smooth muscle myosin tailpieces differentially affect filament assembly, and suggest that homogeneous thick filaments containing SM1 or SM2 myosin could serve distinct functions within smooth muscle cells.
Rovner
Arthur S
AS
Department of Molecular Physiology and Biophysics, University of Vermont, Burlington, VT 05405, USA. rovner@physiology.med.uvm.edu
Fagnant
Patricia M
PM
Lowey
Susan
S
Trybus
Kathleen M
KM
D008841:Actin Cytoskeleton / Q000737:chemistry / Q000378:metabolism / Q000648:ultrastructure; D017398:Alternative Splicing; D000595:Amino Acid Sequence; D000818:Animals; D002645:Chickens; D003460:Crystallization; D019281:Dimerization; D005895:Gizzard, Avian; D008854:Microscopy, Electron; D008969:Molecular Sequence Data; D018995:Myosin Heavy Chains / Q000737:chemistry* / Q000378:metabolism / Q000648:ultrastructure*; D020033:Protein Isoforms / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D020836:Protein Structure, Quaternary; D011817:Rabbits; D051381:Rats; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism / Q000648:ultrastructure; D024745:Smooth Muscle Myosins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism / Q000648:ultrastructure*; D012995:Solubility
D020033:Protein Isoforms; D011994:Recombinant Proteins; D024745:Smooth Muscle Myosins; D018995:Myosin Heavy Chains