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DNA-binding sequence specificity of DUX4.

Yu Zhang1, John K Lee2, Erik A Toso1

  • 1Lillehei Heart Institute, University of Minnesota, Minneapolis, MN 55455 USA ; Department of Pediatrics, University of Minnesota, Minneapolis, MN 55455 USA.

Skeletal Muscle
|January 30, 2016
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Summary
This summary is machine-generated.

DUX4 binding requires two specific TAAT motifs for maximal activity, with variations impacting DNA binding and gene activation. This research clarifies DUX4

Keywords:
DUX4FSHDFacioscapulohumeral muscular dystrophySELEX

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Area of Science:

  • Molecular Biology
  • Genetics
  • Biochemistry

Background:

  • Misexpression of the double homeodomain transcription factor DUX4 is linked to facioscapulohumeral muscular dystrophy (FSHD).
  • A DNA-binding consensus for DUX4 with two TAAT motifs was identified, but variations and their effects remain unclear.
  • The DUX4 binding site in the Pitx1 promoter differs from the consensus, raising questions about its role as a target.

Purpose of the Study:

  • To determine the precise DNA-binding specificity of DUX4.
  • To investigate DUX4's tolerance to sequence variations within its binding motifs.
  • To assess the transcriptional activation potential of different DUX4 binding site configurations.

Main Methods:

  • Utilized unbiased and consensus sequence-driven approaches to map DUX4 DNA-binding specificity.
  • Performed DNA-binding assays and transcriptional activation studies.
  • Investigated the impact of DUX4 binding site number and arrangement on transcriptional output.

Main Results:

  • Optimal DUX4 binding and transcriptional activation occur with two TAAT motifs separated by a cytosine (C).
  • DUX4 prefers thymine (T) at the second TAAT motif's variable position and shows no activity on single 'half-sites'.
  • DUX4 does not bind the Pitx1 promoter sequence, indicating it is not a direct DUX4 target, and demonstrates synergistic activation with multiple binding sites.

Conclusions:

  • These findings elucidate the specific DNA sequence requirements for DUX4 binding and transcriptional activity.
  • The study clarifies DUX4's sequence preferences and highlights the importance of multiple binding sites for robust gene activation.
  • The results challenge the role of PITX1 as a direct DUX4 target gene.