Detection of beta-globin mRNA precursor in heterogeneous nuclear ribonucleoprotein associated with U1-RNP by using anti-RNP antibody.
Heterogeneous nuclear RNA-ribonucleoprotein (hnRNP) fractions were isolated from Friend erythroleukemia cells and separated by 15-45% sucrose gradient centrifugation. The distribution of small nuclear RNAs (snRNAs) in hnRNP fractions indicated that the snRNAs are associated with hnRNP particles. HnRNP fractions were incubated with normal IgG or anti-U1 RNP IgG, and the resulting immunocomplexes were isolated by binding to a protein A-Sepharose column. HnRNP was found in bound fractions only when anti-U1 RNP IgG was used. By Northern hybridization of RNA extracted from the immunocomplexes with a beta-globin genomic DNA probe, 15S beta-globin mRNA precursors and 10S mature mRNA were detected. These findings suggest the existence of a complex of U1 RNP particles and hnRNP particles containing beta-globin pre-mRNA.
D000818:Animals; D000918:Antibody Specificity; D001323:Autoantibodies / Q000302:isolation & purification; D002460:Cell Line; D002467:Cell Nucleus / Q000032:analysis; D002499:Centrifugation, Density Gradient; D002846:Chromatography, Affinity; D005622:Friend murine leukemia virus; D005914:Globins / Q000235:genetics*; D034441:Heterogeneous-Nuclear Ribonucleoproteins; D007074:Immunoglobulin G / Q000302:isolation & purification; D004915:Leukemia, Erythroblastic, Acute / Q000378:metabolism; D007942:Leukemia, Experimental / Q000378:metabolism; D051379:Mice; D009693:Nucleic Acid Hybridization; D009694:Nucleic Acid Precursors / Q000032:analysis*; D012322:RNA Precursors; D012326:RNA Splicing; D012333:RNA, Messenger / Q000032:analysis*; D012261:Ribonucleoproteins / Q000032:analysis* / Q000276:immunology
D001323:Autoantibodies; D034441:Heterogeneous-Nuclear Ribonucleoproteins; D007074:Immunoglobulin G; D009694:Nucleic Acid Precursors; D012322:RNA Precursors; D012333:RNA, Messenger; D012261:Ribonucleoproteins; D005914:Globins