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First published online 21 May 2008
doi: 10.1242/jcs.027250

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RNA polymerase II activity is located on the surface of protein-rich transcription factories

Christopher H. Eskiw^*, Alexander Rapp, David R. F. Carter and Peter R. Cook

Sir William Dunn School of Pathology, University of Oxford, South Parks Road, Oxford, OX1 3RE, UK

Author for correspondence (e-mail: peter.cook{at}path.ox.ac.uk)

Accepted 7 April 2008

We used electron spectroscopic imaging to map nucleoplasmic transcription sites in human cells at unprecedented resolution. HeLa cells were permeabilised, nascent transcripts were extended in BrUTP by 40 nucleotides and the resulting BrRNA immunolabelled with gold particles before structures were viewed. Nascent RNA is almost invariably associated with polymorphic and nitrogen-rich (but phosphorus-poor) structures with a diameter of 87 nm and mass of 10 MDa (calculated by reference to nucleosomes with known numbers of phosphorus and nitrogen atoms). Structures with similar atomic signatures and diameters were observed using correlative microscopy and in unpermeabilised cells. Our results are consistent with RNA synthesis occurring on the surface of these huge protein-rich transcription factories.

Key words: Bromo-uridine triphosphate, Electron microscopy, Electron spectroscopic imaging, Transcription