The proteomes of transcription factories containing RNA polymerases I, II or III.
Nat Methods
; 8(11): 963-8, 2011 Sep 25.
Article
em En
| MEDLINE
| ID: mdl-21946667
ABSTRACT
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.
Texto completo:
1
Base de dados:
MEDLINE
Assunto principal:
Transcrição Gênica
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RNA Polimerases Dirigidas por DNA
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Proteoma
Limite:
Humans
Idioma:
En
Ano de publicação:
2011
Tipo de documento:
Article