Splicing Speckles Are Not Reservoirs of RNA Polymerase II, but Contain an Inactive Form, Phosphorylated on Serine2 Residues of the C-Terminal Domain

SQ Xie, S Martin, PV Guillot, DL Bentley… - Molecular biology of …, 2006 - Am Soc Cell Biol
SQ Xie, S Martin, PV Guillot, DL Bentley, A Pombo
Molecular biology of the cell, 2006Am Soc Cell Biol
“Splicing speckles” are major nuclear domains rich in components of the splicing machinery
and polyA+ RNA. Although speckles contain little detectable transcriptional activity, they are
found preferentially associated with specific mRNA-coding genes and gene-rich R bands,
and they accumulate some unspliced pre-mRNAs. RNA polymerase II transcribes mRNAs
and is required for splicing, with some reports suggesting that the inactive complexes are
stored in splicing speckles. Using ultrathin cryosections to improve optical resolution and …
“Splicing speckles” are major nuclear domains rich in components of the splicing machinery and polyA+ RNA. Although speckles contain little detectable transcriptional activity, they are found preferentially associated with specific mRNA-coding genes and gene-rich R bands, and they accumulate some unspliced pre-mRNAs. RNA polymerase II transcribes mRNAs and is required for splicing, with some reports suggesting that the inactive complexes are stored in splicing speckles. Using ultrathin cryosections to improve optical resolution and preserve nuclear structure, we find that all forms of polymerase II are present, but not enriched, within speckles. Inhibition of polymerase activity shows that speckles do not act as major storage sites for inactive polymerase II complexes but that they contain a stable pool of polymerase II phosphorylated on serine2 residues of the C-terminal domain, which is transcriptionally inactive and may have roles in spliceosome assembly or posttranscriptional splicing of pre-mRNAs. Paraspeckle domains lie adjacent to speckles, but little is known about their protein content or putative roles in the expression of the speckle-associated genes. We find that paraspeckles are transcriptionally inactive but contain polymerase II, which remains stably associated upon transcriptional inhibition, when paraspeckles reorganize around nucleoli in the form of caps.
Am Soc Cell Biol