Details

Autor(en) / Beteiligte

• Melnik, Svitlana
• Deng, Binwei
• Papantonis, Argyris
• Baboo, Sabyasachi
• Carr, Ian M
• Cook, Peter R

Titel

The proteomes of transcription factories containing RNA polymerases I, II or III

Ist Teil von

• Nature methods, 2011-11, Vol.8 (11), p.963-968

Ort / Verlag

United States

Erscheinungsjahr

2011

Quelle

MEDLINE

Beschreibungen/Notizen

• Human nuclei contain three RNA polymerases (I, II and III) that transcribe different groups of genes; the active forms of all three are difficult to isolate because they are bound to the substructure. Here we describe a purification approach for isolating active RNA polymerase complexes from mammalian cells. After isolation, we analyzed their protein content by mass spectrometry. Each complex represents part of the core of a transcription factory. For example, the RNA polymerase II complex contains subunits unique to RNA polymerase II plus various transcription factors but shares a number of ribonucleoproteins with the other polymerase complexes; it is also rich in polymerase II transcripts. We also describe a native chromosome conformation capture method to confirm that the complexes remain attached to the same pairs of DNA templates found in vivo.