title: Structural effect of the L16Q, K50E, and R53P mutations on homeodomain of pituitary homeobox protein 2.
abstract: The transcription factor pituitary homeobox protein 2 (PITX2) is involved in genetic control of development. Mutations in PITX2, most in the homeodomain, cause the autosomal-dominant disorder Rieger syndrome. The mutants L16Q, K50E and R53P destabilize the structure and disrupt DNA-binding activity. The biological functions of these mutants have been characterized but not the structural basis behind the loss of DNA-binding activity. We performed multiple molecular dynamics simulations at 37°C to investigate the structural and dynamic effects of the 3 PITX2 homeodomain mutants. Compared with the wild type (WT), the L16Q mutant induces a kink in the α3 helix, which is stabilized by the hydrogen bond of Q21-R59. The disruption in backbone hydrogen bonds of V47-N51 and W48-R52 leads to a kink formation in the α3 helix of K50E. The R53P mutant alters the relative orientation of helices, which is apparently stabilized by the formation of new hydrogen bonds of T38-Q11, T38-Q12, T38-R2, N39-R2, L40-Q1, L40-R2, and T41-Q4. The hydrophobic core residues F8, L13, L40 and V45 change their positions in all mutants to break the hydrophobic core. Thus, changes in helical orientations and hydrophobic core cause rearrangement of the DNA-binding surface and disrupt DNA-binding activity in the mutants. The structural and molecular dynamics properties of 3 PITX2 homeodomain mutants differ from those of the WT, especially in formation of a kink in the recognition helix, change in the packing of helices and disruption of the hydrophobic core. This structural basis for the loss of DNA-binding activity for these polymorphisms may help in understanding the effect of mutations on other homeodomains with other diseases.
authors.affiliation: Institute of Biomedical Sciences, Academia Sinica, Taipei 115, Taiwan, ROC.
authors.forename: M
authors.initials: M
authors.lastname: Rajasekaran
authors.forename: Chinpan
authors.initials: C
authors.lastname: Chen
mesh_terms: D019943:Amino Acid Substitution; D004268:DNA-Binding Proteins / Q000737:chemistry / Q000235:genetics; D018398:Homeodomain Proteins / Q000737:chemistry* / Q000235:genetics*; D057927:Hydrophobic and Hydrophilic Interactions; D056004:Molecular Dynamics Simulation; D009154:Mutation; D011487:Protein Conformation; D054730:Protein Interaction Domains and Motifs / Q000235:genetics*; D017433:Protein Structure, Secondary; D055672:Static Electricity; D014157:Transcription Factors / Q000737:chemistry* / Q000235:genetics*; D000097577:Homeobox Protein PITX2
chemical_list: D004268:DNA-Binding Proteins; D018398:Homeodomain Proteins; D014157:Transcription Factors; D000097577:Homeobox Protein PITX2