Unraveling the mechanism of a potent transcriptional activator.
Despite their enormous potential as novel research tools and therapeutic agents, artificial transcription factors (ATFs) that up-regulate transcription robustly in vivo remain elusive. In investigating an ATF that does function exceptionally well in vivo, we uncovered an unexpected relationship between transcription function and a binding interaction between the activation domain and an adjacent region of the DNA binding domain. Disruption of this interaction leads to complete loss of function in vivo, even though the activation domain is still able to bind to its target in the transcriptional machinery. We propose that this interaction parallels those between natural activation domains and their regulatory proteins, concealing the activation domain from solvent and the cellular milieu until it binds to its transcriptional machinery target. Inclusion of this property in the future design of ATFs should enhance their efficacy in vivo.
Lu
Zhen
Z
Department of Biochemistry, University of Wisconsin, Madison, 53706, USA.
Rowe
Steven P
SP
Brennan
Brian B
BB
Davis
Sarah E
SE
Metzler
Renee E
RE
Nau
Johnathan J
JJ
Majmudar
Chinmay Y
CY
Mapp
Anna K
AK
Ansari
Aseem Z
AZ
D000595:Amino Acid Sequence; D001665:Binding Sites; D004268:DNA-Binding Proteins; D008969:Molecular Sequence Data; D029701:Saccharomyces cerevisiae Proteins / Q000502:physiology*; D015534:Trans-Activators / Q000737:chemistry / Q000502:physiology*; D014157:Transcription Factors / Q000502:physiology*
D004268:DNA-Binding Proteins; C052130:GAL4 protein, S cerevisiae; D029701:Saccharomyces cerevisiae Proteins; D015534:Trans-Activators; D014157:Transcription Factors