title: LIM domain binding proteins 1 and 2 have different oligomeric states.
abstract: LIM domain binding (Ldb) proteins are important regulators of LIM homeodomain and LIM-only proteins that specify cell fate in many different tissues. An essential feature of these proteins is the ability to self-associate, but there have been no studies that characterise the nature of this self-association. We have used deletion mutagenesis with yeast two-hybrid analysis to define the minimal self-association domains of Ldb1 and Ldb2 as residues 14-200 and 21-197, respectively. We then used a range of different biophysical methods, including sedimentation equilibrium and small-angle X-ray scattering to show that Ldb1(14-200) forms a trimer and Ldb2(21-197) undergoes a monomer-tetramer-octamer equilibrium, where the association in each case is of moderate affinity ( approximately 10(5) M(-1)). These modes of association represent a clear physical difference between these two proteins that otherwise appear to have very similar properties. The levels of association are more complex than previously assumed and emphasise roles of avidity and DNA looping in transcriptional regulation by Ldb1/LIM protein complexes. The abilities of Ldb1 and Ldb2 to form trimers and higher oligomers, respectively, should be considered in models of transcriptional regulation by Ldb1-containing complexes in a wide range of biological processes.
authors.affiliation: School of Molecular Bioscience, Building G08, The University of Sydney, NSW 2006, Australia.
authors.forename: Arwen J
authors.initials: AJ
authors.lastname: Cross
authors.forename: Cy M
authors.initials: CM
authors.lastname: Jeffries
authors.forename: Jill
authors.initials: J
authors.lastname: Trewhella
authors.forename: Jacqueline M
authors.initials: JM
authors.lastname: Matthews
mesh_terms: D000818:Animals; D001665:Binding Sites; D004268:DNA-Binding Proteins / Q000737:chemistry* / Q000378:metabolism; D060588:LIM Domain Proteins; D051379:Mice; D014157:Transcription Factors / Q000737:chemistry* / Q000378:metabolism; D020798:Two-Hybrid System Techniques
chemical_list: D004268:DNA-Binding Proteins; D060588:LIM Domain Proteins; C102084:Ldb1 protein, mouse; C106649:Ldb2 protein, mouse; D014157:Transcription Factors