Evidence that all SC-35 domains contain mRNAs and that transcripts can be structurally constrained within these domains.
A fundamental question of mRNA metabolism concerns the spatial organization of the steps involved in generating mature transcripts and their relationship to SC-35 domains, nuclear compartments enriched in mRNA metabolic factors and poly A+ RNA. Because poly A+ RNA in SC-35 domains remains after transcription inhibition, a prevailing view has been that most or all SC-35 domains do not contain protein-encoding mRNAs but stable RNAs with nuclear functions and thus that these compartments do not have direct roles in mRNA synthesis or transport. However, the transcription, splicing, and transport of transcripts from a specific gene have been shown to occur in association with two of these 15-30 nuclear compartments. Here we show that virtually all SC-35 domains can contain specific mRNAs and that these persist in SC-35 domains after treatment with three different transcription-inhibitory drugs. This suggests perturbation of an mRNA transport step that normally occurs in SC-35 domains and is post-transcriptional but still dependent on ongoing transcription. Finally, even after several hours of transcription arrest, these transcripts do not disperse from SC-35 domains, indicating that they are structurally constrained within them. Our findings importantly suggest a spatially direct role for all SC-35 domains in the coupled steps of mRNA metabolism and transport.
Shopland
Lindsay S
LS
Department of Cell Biology, University of Massachusetts Medical School, 55 Lake Avenue North (S3-138), Worcester, MA 01655-0002, USA.
Johnson
Carol V
CV
Lawrence
Jeanne B
JB
D002460:Cell Line; D002467:Cell Nucleus / Q000378:metabolism; D002843:Chromatin / Q000378:metabolism; D003094:Collagen / Q000096:biosynthesis; D024042:Collagen Type I / Q000096:biosynthesis; D000090602:Collagen Type I, alpha 1 Chain; D018076:DNA, Complementary / Q000378:metabolism; D006801:Humans; D017404:In Situ Hybridization, Fluorescence; D008856:Microscopy, Fluorescence; D009693:Nucleic Acid Hybridization; D017434:Protein Structure, Tertiary; D012333:RNA, Messenger / Q000378:metabolism*; D017411:Ribonucleoproteins, Small Nuclear / Q000378:metabolism; D014158:Transcription, Genetic
D002843:Chromatin; D024042:Collagen Type I; D000090602:Collagen Type I, alpha 1 Chain; D018076:DNA, Complementary; D012333:RNA, Messenger; D017411:Ribonucleoproteins, Small Nuclear; D003094:Collagen