Cytoplasmic assembly of small nuclear ribonucleoprotein particles from 6 S and 20 S RNA-free intermediates in L929 mouse fibroblasts.
Newly transcribed small nuclear RNAs (snRNAs) appear transiently in the cytoplasm where they assemble with snRNP core proteins (B, D, E, F, and G) stored in large pools of snRNA-free intermediates before returning permanently to the nucleus. In this report, the cytoplasmic assembly of snRNP core particles in L929 mouse fibroblasts was investigated by kinetic analysis of assembly intermediates resolved on sucrose gradients. Immunoprecipitation of gradient fractions with anti-snRNP autoimmune antisera identify pools of 6 and 20 S snRNA-free snRNP protein intermediates. The snRNP B protein has a heterodisperse sedimentation from 4 to 20 S with peaks at 6 and 20 S, and the snRNP D protein is in a bimodal distribution at 6 and 20 S. At 6 S the D protein is assembled with the E, F, and G proteins into a RNA-free core particle with a stoichiometry of D4EFG. SnRNP assembly proceeds by snRNA assembling initially with the 6 S D4EFG particle and then two copies of the B protein to form an 11-15 S SnRNP particle. The 20 S forms of the D protein in the cytoplasm are less stable than the 6 S D4EFG particle. The U1-specific A and C proteins leak from isolated nuclei and appear in the cytoplasmic fractions where they sediment from 10 to 20 S and from 4 to 8 S, respectively.
Department of Anatomical Sciences State University of New York, Stony Brook 11794.
R A
RA
Sauterer
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A
Goyal
G W
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Zieve
D000818:Animals; D015153:Blotting, Western; D002458:Cell Fractionation; D004591:Electrophoresis, Polyacrylamide Gel; D005347:Fibroblasts / Q000378:metabolism; D007700:Kinetics; D007739:L Cells; D051379:Mice; D011233:Precipitin Tests; D012313:RNA / Q000235:genetics*; D012322:RNA Precursors / Q000235:genetics; D012261:Ribonucleoproteins / Q000235:genetics*; D017411:Ribonucleoproteins, Small Nuclear; D014158:Transcription, Genetic
D012322:RNA Precursors; D012261:Ribonucleoproteins; D017411:Ribonucleoproteins, Small Nuclear; D012313:RNA