The cytoplasmic sites of the snRNP protein complexes are punctate structures that are responsive to changes in metabolism and intracellular architecture.
Five anti-Sm monoclonal antibodies, Y12, 7.13, KSm4, KSm6, and 128, stain similar discrete punctate structures distributed throughout the cytoplasm of hamster fibroblasts in addition to the expected intense nuclear staining. Several criteria suggest the cytoplasmic staining reflects the cytoplasmic pools of snRNP core proteins. The relative intensity of the cytoplasmic staining is similar to the 30% relative abundance of the cytoplasmic snRNP core proteins compared to the nuclear snRNP core proteins based on cell-fractionation studies. Moreover, the cytoplasmic staining is removed by the same extraction conditions that solubilize the pools of cytoplasmic snRNP core proteins. The cytoplasmic sites of staining are typically spherical but heterogeneous in diameter (0.2-0.5 microm). The larger particles greatly exceed the diameter of individual snRNP core particles and are likely to represent centers of many snRNP proteins or snRNP protein complexes. The staining, though punctate, is evenly dispersed throughout the cytoplasm with no evidence of major compartmentalization. The cytoplasmic staining pattern collapses into larger foci of intensely staining structures when cellular energy levels are depleted or when cells are exposed to hypertonic medium. Unlike the normal sites of snRNP protein cytoplasmic staining, these larger collapsed foci resist detergent extraction. These results suggest that the cytoplasmic staining identified with the anti-Sm monoclonal antibodies represents the large pools of snRNP core proteins in the cytoplasm.
Zieve
G W
GW
Department of Pathology, SUNY Stony Brook, Stony Brook, New York, 11794-8691, USA. GZIEVE@PATH.SOM.SUNYSB.EDU
D000818:Animals; D000911:Antibodies, Monoclonal / Q000032:analysis; D001323:Autoantibodies / Q000032:analysis; D001324:Autoantigens / Q000276:immunology; D002478:Cells, Cultured; D006224:Cricetinae; D003470:Culture Media; D003593:Cytoplasm / Q000737:chemistry* / Q000378:metabolism*; D004734:Energy Metabolism; D005347:Fibroblasts; D006982:Hypertonic Solutions; D046911:Macromolecular Substances; D017411:Ribonucleoproteins, Small Nuclear / Q000737:chemistry* / Q000378:metabolism*; D013194:Staining and Labeling; D055517:snRNP Core Proteins
D000911:Antibodies, Monoclonal; D001323:Autoantibodies; D001324:Autoantigens; D003470:Culture Media; D006982:Hypertonic Solutions; D046911:Macromolecular Substances; D017411:Ribonucleoproteins, Small Nuclear; D055517:snRNP Core Proteins