The RNA polymerase II transcription factor TFIIF is a heterodimer with 2 subunits: one, referred to as RAP74, of approximately 70 kD and the other, known as RAP30 or GTF2F2, of 30 kD.
TFIIF affects RNA polymerase II activity both during the initiation and elongation stages of RNA transcription. TFIIF appears to assist TFIIB in recruiting RNA polymerase II into a preinitiation complex. It also affects RNA transcription by suppressing transient pausing of the polymerase at locations on the DNA template.
TFIIF mediates the association of RNA polymerase II with promoter sequences containing transcription factors IID, IIB, and IIA (DAB complex). Cloned human RAP 30 was sufficient for the recruitment of RNA polymerase II to the DAB complex. This ability of RAP 30 to recruit RNA polymerase to a promoter is also a characteristic of sigma factors in prokaryotes.
Chromatin was prepared from Hela cells according to the Abcam X-ChIP protocol. Cells were fixed with formaldehyde for 10mins. The ChIP was performed with 25µg of chromatin, 8µg of ab28179 (blue), and 20µl of Protein A/G sepharose beads. No antibody was added to the beads control (yellow). The immunoprecipitated DNA was quantified by real time PCR (Taqman and sybr green approach). Primers and probes for the actively transcribed genes GAPDH and ACT1 are located in the core promoter.
Anti-TFIIF antibody (ab28179)参考文献
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