Both the TATA and CCAAT boxes are widespread promoter elements and their binding proteins, TBP and NF-Y, are extremely conserved in evolution, NF-Y is composed os three subunits, NF-YA, NF-YB and NF-YC, all necessary for DNA binding. NF-YB and NF-YC contain a putative histone-like motif, a domain also present in TBP-associated factors (TAF(II)s) and in the subunits of the transcriptional repressor NC2. Immunopurification of holo-TFIID with anti-TBP and anti-TAF(II)100 antibodies indicates that a fraction of NF-YB associates with TFIID in the absence of NF-YA. Sedimentation velocity centrifugation experiments confirm that two pools of NF-YB, and most likely NF-YC, exist: one associated with NF-YA and binding to the CCAAT box; another involved in high molecular weight complexes. We started to dissect NF-Y-TFIID interactions by showing that: (i) NF-YB and NF-YC interact with TBP in solution, both separately and once bound to each other; (ii) short stretches of both NF-YB and NF-YC located within the evolutionary conserved domains, adjacent to the putative histone fold motifs, are necessary for TBP binding; (iii) TBP single amino acid mutants in the HS2 helix, previously shown to be defective in NC2 binding, are also unable to bind NF-YB and NF-YC.
|Titolo:||CCAAT binding NF-Y-TBP interactions : NF-YB and NF-YC require short domains adjacent to their histone fold motifs for association with TBP basic residues|
MANTOVANI, ROBERTO (Ultimo)
|Parole Chiave:||Histones ; Protein Folding ; TATA Box ; Amino Acid Sequence ; Animals ; Binding Sites ; CCAAT-Enhancer-Binding Proteins ; Centrifugation, Density Gradient ; DNA-Binding Proteins ; Escherichia coli ; Humans ; Mice ; Molecular Sequence Data ; Molecular Weight ; Rats ; TATA-Box Binding Protein ; Transcription Factor TFIID ; Transcription Factors ; Transcription Factors, TFII|
|Settore Scientifico Disciplinare:||Settore BIO/18 - Genetica|
|Data di pubblicazione:||1-giu-1997|
|Digital Object Identifier (DOI):||10.1093/nar/25.11.2174|
|Appare nelle tipologie:||01 - Articolo su periodico|