title: Mutagenesis of the Runt domain defines two energetic hot spots for heterodimerization with the core binding factor beta subunit.
abstract: Core-binding factors (CBFs) are a small family of heterodimeric transcription factors that play critical roles in several developmental pathways and in human disease. Mutations in CBF genes are found in leukemias, bone disorders, and gastric cancers. CBFs consist of a DNA-binding CBF alpha subunit (Runx1, Runx2, or Runx3) and a non-DNA-binding CBF beta subunit. CBF alpha binds DNA in a sequence-specific manner, whereas CBF beta enhances DNA binding by CBF alpha. Both DNA binding and heterodimerization with CBF beta are mediated by a single domain in the CBF alpha subunits known as the "Runt domain." We analyzed the energetic contribution of amino acids in the Runx1 Runt domain to heterodimerization with CBF beta. We identified two energetic "hot spots" that were also found in a similar analysis of CBF beta (Tang, Y.-Y., Shi, J., Zhang, L., Davis, A., Bravo, J., Warren, A. J., Speck, N. A., and Bushweller, J. H. (2000) J. Biol. Chem. 275, 39579-39588). The importance of the hot spot residues for Runx1 function was demonstrated in in vivo transient transfection assays. These data refine the structural analyses and further our understanding of the Runx1-CBF beta interface.
authors.affiliation: Department of Biochemistry, Dartmouth Medical School, Hanover, New Hampshire 03755, USA.
authors.forename: Lina
authors.initials: L
authors.lastname: Zhang
authors.forename: Zhe
authors.initials: Z
authors.lastname: Li
authors.forename: Jiangli
authors.initials: J
authors.lastname: Yan
authors.forename: Padmanava
authors.initials: P
authors.lastname: Pradhan
authors.forename: Takeshi
authors.initials: T
authors.lastname: Corpora
authors.forename: Matthew D
authors.initials: MD
authors.lastname: Cheney
authors.forename: Jerónimo
authors.initials: J
authors.lastname: Bravo
authors.forename: Alan J
authors.initials: AJ
authors.lastname: Warren
authors.forename: John H
authors.initials: JH
authors.lastname: Bushweller
authors.forename: Nancy A
authors.initials: NA
authors.lastname: Speck
mesh_terms: D000409:Alanine / Q000737:chemistry; D000818:Animals; D050659:Core Binding Factor Alpha 1 Subunit; D050676:Core Binding Factor Alpha 2 Subunit; D050657:Core Binding Factor alpha Subunits; D018076:DNA, Complementary / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000737:chemistry* / Q000235:genetics* / Q000378:metabolism; D019281:Dimerization; D004305:Dose-Response Relationship, Drug; D029721:Drosophila Proteins; D007700:Kinetics; D008156:Luciferases / Q000378:metabolism; D009682:Magnetic Resonance Spectroscopy; D051379:Mice; D008856:Microscopy, Fluorescence; D008954:Models, Biological; D008958:Models, Molecular; D016297:Mutagenesis, Site-Directed; D009687:Nuclear Proteins; D010957:Plasmids / Q000378:metabolism; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D011518:Proto-Oncogene Proteins; D013050:Spectrometry, Fluorescence; D050656:Transcription Factor AP-2; D014157:Transcription Factors / Q000737:chemistry* / Q000235:genetics*; D014162:Transfection; D014508:Urea / Q000494:pharmacology
chemical_list: D050659:Core Binding Factor Alpha 1 Subunit; D050676:Core Binding Factor Alpha 2 Subunit; D050657:Core Binding Factor alpha Subunits; D018076:DNA, Complementary; D004268:DNA-Binding Proteins; D029721:Drosophila Proteins; D009687:Nuclear Proteins; D011518:Proto-Oncogene Proteins; C493726:Runx1 protein, mouse; C493716:Runx2 protein, mouse; D050656:Transcription Factor AP-2; D014157:Transcription Factors; C080833:run protein, Drosophila; D014508:Urea; D008156:Luciferases; D000409:Alanine