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      Observation of unphosphorylated STAT3 core protein binding to target dsDNA by PEMSA and X-ray crystallography.

      Febs Letters
      Animals, Binding Sites, Binding, Competitive, Circular Dichroism, Crystallography, X-Ray, DNA, chemistry, genetics, metabolism, Electrophoretic Mobility Shift Assay, methods, Humans, Luminescent Proteins, Models, Molecular, Nucleic Acid Conformation, Phosphopeptides, Phosphorylation, Protein Binding, Protein Structure, Secondary, Protein Structure, Tertiary, STAT3 Transcription Factor, Tyrosine

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          Abstract

          The STAT3 transcription factor plays a central role in a wide range of cancer types where it is over-expressed. Previously, phosphorylation of this protein was thought to be a prerequisite for direct binding to DNA. However, we have now shown complete binding of a purified unphosphorylated STAT3 (uSTAT3) core directly to M67 DNA, the high affinity STAT3 target DNA sequence, by a protein electrophoretic mobility shift assay (PEMSA). Binding to M67 DNA was inhibited by addition of increasing concentrations of a phosphotyrosyl peptide. X-ray crystallography demonstrates one mode of binding that is similar to that known for the STAT3 core phosphorylated at Y705. Copyright © 2013 Federation of European Biochemical Societies. Published by Elsevier B.V. All rights reserved.

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