title: Measuring specific interaction of transcription factor ZmDREB1A with its DNA responsive element at the molecular level.
abstract: Specific interactions between transcription factors and DNA responsive elements are of fundamental importance in understanding how genetic regulatory proteins control gene transcription. Here we have developed a new method of using atomic force microscopy (AFM) to quantitatively study the single molecular specific interaction between ZmDREB1A, a transcription factor from maize, and its DNA responsive element, dehydration-responsive element (DRE) with core sequence A/GCCGAC. It was found that ZmDREB1A bound to both DRE ACCGAC and GCCGAC efficiently. The single molecular interaction forces of ZmDREB1A with DRE A/GCCGAC were determined to be 101 +/- 5 and 108 +/- 3 pN, respectively. The point mutation of ZmDREB1A in its DNA-binding domain or single base substitution of the DRE core sequence greatly reduced the binding affinity, demonstrating the high sensitivity of the AFM measurements. AFM is expected to be a simple, quick, sensitive and reliable method that offers valuable information for the characterization of transcription factors and the identification of their potential DNA responsive elements in functional genomics research.
authors.affiliation: Institute of Chemistry, Chinese Academy of Sciences, Beijing 100080, China.
authors.forename: Yaxin
authors.initials: Y
authors.lastname: Jiang
authors.forename: Feng
authors.initials: F
authors.lastname: Qin
authors.forename: Yiqin
authors.initials: Y
authors.lastname: Li
authors.forename: Xiaohong
authors.initials: X
authors.lastname: Fang
authors.forename: Chunli
authors.initials: C
authors.lastname: Bai
mesh_terms: D029681:Arabidopsis Proteins; D001483:Base Sequence; D001665:Binding Sites; D018625:Microscopy, Atomic Force; D008969:Molecular Sequence Data; D009154:Mutation; D010937:Plant Growth Regulators / Q000502:physiology; D020218:Response Elements; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism*; D003313:Zea mays / Q000235:genetics
chemical_list: D029681:Arabidopsis Proteins; C114578:DREB1A protein, Arabidopsis; D010937:Plant Growth Regulators; D014157:Transcription Factors