Conservative Val⁴⁷ residue of POU homeodomain: role in DNA recognition

Abstract 

Conservative Val⁴⁷ residue, located in the third recognition helix of the Oct-2 POU domain, was alternately substituted with other 19 amino acids. Affinity and specificity of interaction with oct-site ATGCAAANGA and homeo-specific site ATAANGA were determined for all mutants. The wild type protein (with Val⁴⁷) has maximal affinity and specificity in POU domain interaction with octamer sequence. However, V47I mutant showed stronger interaction with homeo-specific site. The highest specificity of interaction with homeo-site was recorded for V47S mutant. We conclude that only Val⁴⁷ provides sequence-specific high-affinity binding of POU proteins with octamer targets other than the homeo-specific site. It is shown also that damages caused by point mutations may be at least partially compensated by participation in the oct-site recognition of both POUh and POUs domains.

Keywords:  Oct-2 protein , POU domain mutants , Oct-sequence , Homeo-specific site , Protein-DNA recognition