A direct interaction between a glutamine-rich activator and the N terminus of TFIIB can mediate transcriptional activation in vivo.
Studies examining the mechanism by which transcriptional activators function have suggested that the general transcription factor IIB (TFIIB) can be a target for certain regulatory proteins. For example, we showed previously that expression of a mutant form of TFIIB can specifically inhibit activation in vivo mediated by the strong, glutamine-rich activator protein GAL4-ftzQ. Using transient cotransfection assays, we have defined the regions in both GAL4-ftzQ and TFIIB that are required for activity in vivo and provide evidence that a potential zinc finger structure at the N terminus of TFIIB is necessary for the observed functional interaction between the two proteins. Using a protein binding assay, we have demonstrated that GAL4-ftzQ can specifically interact with TFIIB in vitro. This interaction requires the same regions in both molecules necessary for function in vivo and is reduced or eliminated by mutations predicted to disrupt the zinc finger in TFIIB. These results support the idea that a direct interaction between a regulatory protein and TFIIB can be important for transcriptional activation in vivo and, combined with previous data of others, suggest that different activators can function by contacting distinct regions of TFIIB.
Colgan
J
J
Department of Biological Sciences, Columbia University, New York, New York 10027.
Ashali
H
H
Manley
J L
JL
D000595:Amino Acid Sequence; D000818:Animals; D002478:Cells, Cultured; D004252:DNA Mutational Analysis; D004330:Drosophila / Q000166:cytology; D029721:Drosophila Proteins; D051717:Fushi Tarazu Transcription Factors; D008969:Molecular Sequence Data; D011485:Protein Binding; D012319:RNA Polymerase II / Q000378:metabolism; D011993:Recombinant Fusion Proteins; D013329:Structure-Activity Relationship; D015534:Trans-Activators / Q000378:metabolism*; D035581:Transcription Factor TFIIB; D014157:Transcription Factors / Q000378:metabolism*; D014158:Transcription, Genetic; D016335:Zinc Fingers
D029721:Drosophila Proteins; D051717:Fushi Tarazu Transcription Factors; C080545:GAL4-ftzQ protein, Drosophila; D011993:Recombinant Fusion Proteins; D015534:Trans-Activators; D035581:Transcription Factor TFIIB; D014157:Transcription Factors; D012319:RNA Polymerase II