The spatial distribution of polyribosomes in 3T3 cells and the associated assembly of proteins into the skeletal framework.
Acridine fluorescence reveals polyribosomes in fibroblasts and Triton-extracted skeletal frameworks; simultaneous phase-contrast images show cellular structure. Polyribosomes appear near nuclei of both intact fibroblasts and skeletal frameworks. Simultaneous autoradiography of cells so examined locates radioactive proteins. After synthesis, most proteins diffuse rapidly through the cytoplasm; intact cells autoradiographed afer a 35S pulse show grains throughout. In sharp contrast, extraction with Triton leaves only radioactive skeletal proteins, which, although released from ribosomes, are near polyribosomes after a pulse. After a chase, skeletal-associated radioactivity is found throughout the framework structure. However, skeletal proteins migrate only if protein synthesis continues. Emetine administered following a pulse block protein migration; skeletal framework radioactivity remains near polyribosomes. This also indicates limited exchange between skeletal framework and soluble cytoplasmic proteins. The fact that proteins insert themselves into the skeletal framework at or near their synthesis site, with limited subsequent exchange, appears to contradict current view of protein self-assembly.
D000199:Actins / Q000096:biosynthesis; D000818:Animals; D001665:Binding Sites; D002460:Cell Line; D003599:Cytoskeleton / Q000378:metabolism*; D051379:Mice; D008854:Microscopy, Electron; D008856:Microscopy, Fluorescence; D011132:Polyribosomes / Q000378:metabolism*; D012995:Solubility; D014404:Tubulin / Q000096:biosynthesis
D000199:Actins; D014404:Tubulin