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Nuclear PtdIns(4,5)P₂ assembles in a mitotically regulated particle involved in pre-mRNA splicing

¹ Molecular Neuropathobiology Laboratory, Imperial Cancer Research Fund, 44 Lincoln’s Inn Fields, London, WC2A 3PX, UK
² Electron Microscopy Unit, Imperial Cancer Research Fund, 44 Lincoln’s Inn Fields, London, WC2A 3PX, UK

*Author for correspondence (e-mail: g.schiavo{at}icrf.icnet.uk)

Accepted April 9, 2001

Phosphoinositide turnover regulates multiple cellular processes. Compared with their well-known cytosolic roles, limited information is available on the functions of nuclear phosphoinositides. Here, we show that phosphatidylinositol(4,5)-bisphosphate (PtdIns(4,5)P₂) stably associates with electron-dense particles within the nucleus that resemble interchromatin granule clusters. These PtdIns(4,5)P₂-containing structures have a distribution which is cell-cycle dependent and contain components of both the transcriptional and pre-mRNA processing machinery, including RNA polymerase II and the splicing factor SC-35. Immunodepletion and add-back experiments demonstrate that PtdIns(4,5)P₂ and associated factors are necessary but not sufficient for pre-mRNA splicing in vitro, indicating a crucial role for PtdIns(4,5)P₂-containing complexes in nuclear pre-mRNA processing.

Key words: Mitosis, Phosphatidylinositol(4,5)-bisphosphate, RNA polymerase II

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