In my proposal I plan to investigate the role of TFIID and novel TAF-containing complexes in the regulation of gene expression and to highlight differences between the distinct complexes in transcription regulation.
The objectives of my proposal are centered around dissection of TFIID, TFTC, STAGA and P/CAF specific role(s) in genes expression regulation and my objectives are the following:
-Identification of TFIID, TFTC, STAGA and P/CAF target genes. Do these complexes regulate different promoters or act on the same promoters with different functions that could be partially redundant or not? Or are different combinations of them required to fulfil gene regulation in the complexity of human genome?
-Dissection of their mechanisms and dynamics in transcription regulation. Promoter's histones acetylation is an early event during gene activation, since three out of four of these complexes contain a histone acetyl transferase enzyme, it is crucial to understand if these activities play the same role in different contexts or if they have different targets and then which is the meaning of target acetylation.
To answer these questions I plan to use the following approach: RNA interference of different subunits: common to all complexes, shared by some but not all of the complexes and complex scientific. The comparison of gene expression profiles, obtained with cDNA arrays, by subtractive analysis of the different samples will permit the creation of categories of genes regulate either by different, or combinations of complexes.
On some of these genes a more detailed analysis will be performed using chromatin immunoprecipitations to elucidate the interplay of these factors with the other components of the transcription machinery. The accomplishment of these objectives will clarify important transcriptional mechanisms and furnish additional knowledge of outstanding interest to the further understanding of cell biology.
Fields of science
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