Coiled bodies without coilin.
Nuclei assembled in vitro in Xenopus egg extract contain coiled bodies that have components from three different RNA processing pathways: pre-mRNA splicing, pre-rRNA processing, and histone pre-mRNA 3'-end formation. In addition, they contain SPH-1, the Xenopus homologue of p80-coilin, a protein characteristic of coiled bodies. To determine whether coilin is an essential structural component of the coiled body, we removed it from the egg extract by immunoprecipitation. We showed that nuclei with bodies morphologically identical to coiled bodies (at the light microscope level) formed in such coilin-depleted extract. As expected, these bodies did not stain with antibodies against coilin. Moreover, they failed to stain with an antibody against the Sm proteins, although Sm proteins associated with snRNAs were still present in the extract. Staining of the coilin- and Sm-depleted coiled bodies was normal with antibodies against two nucleolar proteins, fibrillarin and nucleolin. Similar results were observed when Sm proteins were depleted from egg extract: staining of the coiled bodies with antibodies against the Sm proteins and coilin was markedly reduced but bright nucleolin and fibrillarin staining remained. These immunodepletion experiments demonstrate an interdependence between coilin and Sm snRNPs and suggest that neither is essential for assembly of nucleolar components in coiled bodies. We propose that coiled bodies are structurally heterogeneous organelles in which the components of the three RNA processing pathways may occur in separate compartments.
Bauer
D W
DW
Department of Embryology, Carnegie Institution, Baltimore, Maryland 21210, USA.
Gall
J G
JG
D000818:Animals; D015152:Blotting, Northern; D015153:Blotting, Western; D002457:Cell Extracts / Q000737:chemistry; D002466:Cell Nucleolus / Q000737:chemistry / Q000378:metabolism* / Q000648:ultrastructure; D002467:Cell Nucleus / Q000502:physiology / Q000648:ultrastructure*; D005591:Chemical Fractionation; D002868:Chromosomal Proteins, Non-Histone / Q000378:metabolism; D005260:Female; D005455:Fluorescent Antibody Technique; D008297:Male; D009687:Nuclear Proteins / Q000378:metabolism*; D010063:Ovum / Q000502:physiology*; D010750:Phosphoproteins / Q000378:metabolism; D012326:RNA Splicing; D012342:RNA, Small Nuclear / Q000235:genetics / Q000378:metabolism; D016601:RNA-Binding Proteins; D017411:Ribonucleoproteins, Small Nuclear / Q000235:genetics / Q000378:metabolism*; D013094:Spermatozoa / Q000502:physiology; D029867:Xenopus Proteins; D014982:Xenopus laevis; D000097796:Nucleolin
D002457:Cell Extracts; D002868:Chromosomal Proteins, Non-Histone; D009687:Nuclear Proteins; D010750:Phosphoproteins; D012342:RNA, Small Nuclear; D016601:RNA-Binding Proteins; D017411:Ribonucleoproteins, Small Nuclear; D029867:Xenopus Proteins; D000097796:Nucleolin; C047458:fibrillarin; C068921:p80-coilin