Preliminary three-dimensional model for nematode thick filament core.

Abstract

Understanding the structure and the mechanism of assembly of thick filaments have been long-standing problems in the field of muscle biology. Cores which represent the backbones of thick filaments and consist of paramyosin and associated proteins were isolated from the nematode Caenorhabditis elegans. Electron microscopy of negatively stained and frozen hydrated cores was performed. The resulting images were analyzed by computing their Fourier transforms, three-dimensional reconstruction, and by modeling. A preliminary three-dimensional model is proposed in which the paramyosin constitutes an outer sheath of seven subfilaments about a set of inner 54-nm-long tubules which repeat every 72 nm. The subfilaments are not closely packed but require cross-linking by the internal tubules. Each subfilament consists of two strands of paramyosin molecules which are staggered by 72 nm with respect to one another. This stagger introduces a 22-nm gap between consecutive paramyosin molecules in each strand. An offset of the center of the inner tubules relative to the center of the gap of 6 nm was consistent with the images and their transforms. This model suggests that the nonhelical ends of paramyosin and the unpaired gap between adjacent paramyosin molecules contain sites for the interaction with the inner tubular proteins. The molecular interactions at this locus would appear to be critical in the assembly of thick filaments and their regulation.