The general transcription factor, TFIID, consists of the TATA binding protein (TBP) associated with a series of TBP-associated factors (TAFs) that together participate in the assembly of the transcription preinitiation complex. The largest TFIID component, TAF1 has both acetyltransferase (AT) and kinase activities. We demonstrated previously that TAF1 is necessary for the transcription of an MHC class I gene, and that its intrinsic acetyltransferase activity is essential for both in vitro transcription of naked DNA and in vivo transcription. Of particular interest, MHC class I transcription is inhibited, both in vitro and in vivo, by the transactivator, HIV Tat, which we demonstrated binds to the TAF1 AT domain and inhibits its enzymatic activity. Similarly, TAF7, a 55 kD TFIID component, binds to TAF1 and inhibits its AT activity, resulting in repression of MHC class I transcription. Thus, TAF7 is an intrinsic regulator of transcription. It is released from TFIID , relieving the repression of the TAF1 AT activity and allowing transcription initiation. Our studies determined that after its dissociation from the PIC, TAF7 functionally interacts with both the general transcription factor TFIIH and the elongation factor P-TEFb. Association of TAF7 with TFIIH inhibits its CDK7 kinase activity with the resultant inhibition of TFIIH-mediated phosphorylation of the Pol II CTD Ser-5;binding of TAF7 to the P-TEFb elongation complex inhibits its phosphorylation of Ser-2 of the Pol II CTD. Importantly, we have shown that TAF7 functions in vitro to inhibit transcription at steps after PIC assembly and in vivo co-localizes with P-TEFb and Pol II downstream of the promoter. Thus, in addition to its role in transcription initiation as a TFIID component, TAF7 also functions in the transition from PIC assembly to initiation and elongation. We propose a novel model in which TAF7 regulates the orderly progression of events in transcription, preventing transcription elongation until the steps of transcription initiation are completed and the transcription elongation complex (TEC) is fully assembled. In addition to the regulatory role it plays in TFIID, TAF7 also functions to regulate TAF1-independent transcription. The first evidence for this came from our observation that TAF7 exists in a TFIID-independent form that coelutes with complexes in the 230 Kd range on gel filtration. Interestingly, the kinetics and spectrum of genes affected by siRNA mediated depletion of TAF7 or TAF1 differs between the two. This finding supports the conclusion that TAF7 functions at multiple points in transcription initiation, not just in PIC assembly. The importance of TAF7 in regulating normal transcription is documented by our finding that cells depleted of TAF7, by siRNA or shRNA technology, proliferate poorly. Consistent with the critical role of TAF7 in cell growth, expression profiling reveals that depletion of TAF7 by siRNA results in reduced expression of a large number of genes. To further examine the role of TAF7 in global gene regulation and development we have generated conditional TAF7 knock-out mice. Complete ablation of TAF7 results in embryonic lethality;embryos do not develop beyond d.3-4. Excision of TAF7 in MEFs derived from conditional knock-outs results in aborted cell proliferation and a global reduction in transcription initiation. TAF7 excision at different stages of thymic development differentially affects T cell maturation: lckcre-mediated TAF7 depletion ablates all T cell development whereas depletion of TAF7 at the double positive stage of thymocyte development spares the maturation of single positive cells. The requirement for TAF7 in the periphery is under investigation.
|Gegonne, Anne; Devaiah, Ballachanda N; Singer, Dinah S (2013) TAF7: traffic controller in transcription initiation. Transcription 4:29-33|
|Gegonne, Anne; Tai, Xuguang; Zhang, Jinghui et al. (2012) The general transcription factor TAF7 is essential for embryonic development but not essential for the survival or differentiation of mature T cells. Mol Cell Biol 32:1984-97|
|Lee, Namhoon; Iyer, Shankar S; Mu, Jie et al. (2010) Three novel downstream promoter elements regulate MHC class I promoter activity in mammalian cells. PLoS One 5:e15278|
|Devaiah, Ballachanda N; Lu, Hanxin; Gegonne, Anne et al. (2010) Novel functions for TAF7, a regulator of TAF1-independent transcription. J Biol Chem 285:38772-80|