Identification of carbohydrate binding protein 35 in heterogeneous nuclear ribonucleoprotein complex.
In previous studies, a lectin designated as carbohydrate binding protein 35 (CBP35) was identified in the nucleus and cytoplasm of cultured mouse 3T3 fibroblasts. In the present study, we observed that treatment of Triton X-100 permeabilized 3T3 cells with ribonuclease A released CBP35 from the nuclei, while parallel treatment with deoxyribonuclease I failed to do so. This conclusion was based on (a) immunofluorescence analysis of the nuclear residue after detergent and enzymatic treatments and (b) immunoblotting analysis of the supernatant fraction produced by these treatments. These results indicate that CBP35 may be associated with the ribonucleoprotein elements of the 3T3 cell nuclei. In corroboration with this conclusion, fractionation of the nucleoplasm derived from 3T3 cells on a cesium sulfate gradient (1.25-1.75 g/mL) localized CBP35 in fractions with densities of 1.30-1.32 g/mL, corresponding to the range of densities reported for heterogeneous nuclear ribonucleoprotein complex (hnRNP). Conversely, when nucleoplasm was fractionated on an affinity column of Sepharose derivatized with N-(epsilon-aminocaproyl)-D-galactosamine, the bound and eluted fraction contained RNA, as well as a set of polypeptides whose molecular weights matched those reported for the core particle of hnRNP. One of these polypeptides was identified as CBP35. These results suggest that CBP35 is a component of hnRNP.
D000818:Animals; D002463:Cell Membrane Permeability; D003850:Deoxyribonuclease I / Q000378:metabolism; D005260:Female; D037161:Galectins; D006388:Hemagglutinins / Q000032:analysis*; D034441:Heterogeneous-Nuclear Ribonucleoproteins; D007163:Immunosorbent Techniques; D008970:Molecular Weight; D011817:Rabbits; D012259:Ribonuclease, Pancreatic / Q000378:metabolism; D012261:Ribonucleoproteins / Q000032:analysis*; D014529:Uridine / Q000378:metabolism
D037161:Galectins; D006388:Hemagglutinins; D034441:Heterogeneous-Nuclear Ribonucleoproteins; D012261:Ribonucleoproteins; D003850:Deoxyribonuclease I; D012259:Ribonuclease, Pancreatic; D014529:Uridine