title: Mass spectrometric characterization of sequence-specific complexes of DNA and transcription factor PU.1 DNA binding domain.
abstract: Electrospray ionization mass spectrometry (ESI-MS) has been used to study the noncovalent interaction of the 13.5-kDa DNA binding domain of PU.1 (PU.1-DBD) with specific double-stranded DNA (dsDNA) target molecules. Mixtures of PU.1-DBD protein and wild-type target DNA sequence yielded ESI-MS spectra showing only protein-dsDNA complex ions of 1:1 stoichiometry and free dsDNA. When PU.1-DBD protein, wild type target DNA, and a mutant target DNA lacking the consensus sequence were mixed, only the 1:1 complex with the wild-type DNA was observed, consistent with gel electrophoresis mobility shift assay results, demonstrating the observation of sequence-specific protein-dsDNA complexes using ESI-MS.
authors.affiliation: Environmental Molecular Sciences Laboratory, Pacific Northwest National Laboratory, Richland, Washington, 99352, USA.
authors.forename: X
authors.initials: X
authors.lastname: Cheng
authors.forename: P E
authors.initials: PE
authors.lastname: Morin
authors.forename: A C
authors.initials: AC
authors.lastname: Harms
authors.forename: J E
authors.initials: JE
authors.lastname: Bruce
authors.forename: Y
authors.initials: Y
authors.lastname: Ben-David
authors.forename: R D
authors.initials: RD
authors.lastname: Smith
mesh_terms: D001665:Binding Sites; D004247:DNA / Q000378:metabolism*; D006801:Humans; D013058:Mass Spectrometry; D008969:Molecular Sequence Data; D011485:Protein Binding; D011518:Proto-Oncogene Proteins / Q000378:metabolism*; D015534:Trans-Activators; D000099232:Proto-Oncogene Protein Spi-1
chemical_list: D004247:DNA; D011518:Proto-Oncogene Proteins; D015534:Trans-Activators; D000099232:Proto-Oncogene Protein Spi-1