title: Allostery mediates ligand binding to WWOX tumor suppressor via a conformational switch.
abstract: While being devoid of the ability to recognize ligands itself, the WW2 domain is believed to aid ligand binding to the WW1 domain in the context of a WW1-WW2 tandem module of WW domain-containing oxidoreductase (WWOX) tumor suppressor. In an effort to test the generality of this hypothesis, we have undertaken here a detailed biophysical analysis of the binding of WW domains of WWOX alone and in the context of the WW1-WW2 tandem module to an array of putative proline-proline-x-tyrosine (PPXY) ligands. Our data show that while the WW1 domain of WWOX binds to all ligands in a physiologically relevant manner, the WW2 domain does not. Moreover, ligand binding to the WW1 domain in the context of the WW1-WW2 tandem module is two-to-three-fold stronger than when treated alone. We also provide evidence that the WW domains within the WW1-WW2 tandem module physically associate so as to adopt a fixed spatial orientation relative to each other. Of particular note is the observation that the physical association of the WW2 domain with WW1 blocks access to ligands. Consequently, ligand binding to the WW1 domain not only results in the displacement of the WW2 lid but also disrupts the physical association of WW domains in the liganded conformation. Taken together, our study underscores a key role of allosteric communication in the ability of the WW2 orphan domain to chaperone physiological action of the WW1 domain within the context of the WW1-WW2 tandem module of WWOX.
authors.affiliation: Department of Biochemistry and Molecular Biology, Leonard Miller School of Medicine University of Miami, Miami, FL, 33136, USA.
authors.forename: Brett J
authors.initials: BJ
authors.lastname: Schuchardt
authors.forename: David C
authors.initials: DC
authors.lastname: Mikles
authors.forename: Vikas
authors.initials: V
authors.lastname: Bhat
authors.forename: Caleb B
authors.initials: CB
authors.lastname: McDonald
authors.forename: Marius
authors.initials: M
authors.lastname: Sudol
authors.forename: Amjad
authors.initials: A
authors.lastname: Farooq
mesh_terms: D000495:Allosteric Site; D006801:Humans; D008024:Ligands; D008968:Molecular Conformation; D010088:Oxidoreductases / Q000737:chemistry*; D017434:Protein Structure, Tertiary; D025521:Tumor Suppressor Proteins / Q000737:chemistry*; D000074584:WW Domain-Containing Oxidoreductase
chemical_list: D008024:Ligands; D025521:Tumor Suppressor Proteins; D010088:Oxidoreductases; D000074584:WW Domain-Containing Oxidoreductase; C486071:WWOX protein, human
keywords: WW tandem module; WW-ligand thermodynamics; allosteric communication; binding-coupled dissociation; equilibrium shift