cis Determinants of Promoter Threshold and Activation Timescale.
Although the relationship between DNA cis-regulatory sequences and gene expression has been extensively studied at steady state, how cis-regulatory sequences affect the dynamics of gene induction is not known. The dynamics of gene induction can be described by the promoter activation timescale (AcTime) and amplitude threshold (AmpThr). Combining high-throughput microfluidics with quantitative time-lapse microscopy, we control the activation dynamics of the budding yeast transcription factor, Msn2, and reveal how cis-regulatory motifs in 20 promoter variants of the Msn2-target-gene SIP18 affect AcTime and AmpThr. By modulating Msn2 binding sites, we can decouple AmpThr from AcTime and switch the SIP18 promoter class from high AmpThr and slow AcTime to low AmpThr and either fast or slow AcTime. We present a model that quantitatively explains gene-induction dynamics on the basis of the Msn2-binding-site number, TATA box location, and promoter nucleosome organization. Overall, we elucidate the cis-regulatory logic underlying promoter decoding of TF dynamics.
Department of Chemistry and Chemical Biology, Harvard University, 12 Oxford Street, Cambridge, MA 02138, USA; Howard Hughes Medical Institute, Harvard University, Northwest Laboratory, 52 Oxford Street, Cambridge, MA 02138, USA; Faculty of Arts and Sciences Center for Systems Biology, Harvard University, Northwest Laboratory, 52 Oxford Street, Cambridge, MA 02138, USA.
Anders S
AS
Hansen
Department of Chemistry and Chemical Biology, Harvard University, 12 Oxford Street, Cambridge, MA 02138, USA; Howard Hughes Medical Institute, Harvard University, Northwest Laboratory, 52 Oxford Street, Cambridge, MA 02138, USA; Faculty of Arts and Sciences Center for Systems Biology, Harvard University, Northwest Laboratory, 52 Oxford Street, Cambridge, MA 02138, USA; Department of Molecular and Cellular Biology, Harvard University, Northwest Laboratory, 52 Oxford Street, Cambridge, MA 02138, USA. Electronic address: erin_oshea@harvard.edu.
Erin K
EK
O'Shea
D004268:DNA-Binding Proteins / Q000235:genetics / Q000378:metabolism; D015966:Gene Expression Regulation, Fungal; D012441:Saccharomyces cerevisiae / Q000235:genetics; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics / Q000378:metabolism; D016385:TATA Box; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism; D015533:Transcriptional Activation
D004268:DNA-Binding Proteins; C081935:MSN2 protein, S cerevisiae; D029701:Saccharomyces cerevisiae Proteins; D014157:Transcription Factors