title: Intermolecular disulfide bond influences unphosphorylated STAT3 dimerization and function.
abstract: Signal transducer and activator of transcription 3 (STAT3) is a transcription factor activated by the phosphorylation of tyrosine 705 in response to many cytokines and growth factors. Recently, the roles for unphosphorylated STAT3 (U-STAT3) have been described in response to cytokine stimulation, in cancers, and in the maintenance of heterochromatin stability. It has been reported that U-STAT3 dimerizes, shuttles between the cytoplasm and nucleus, and binds to DNA, thereby driving genes transcription. Although many reports describe the active role of U-STAT3 in oncogenesis in addition to phosphorylated STAT3, the U-STAT3 functional pathway remains elusive.In this report, we describe the molecular mechanism of U-STAT3 dimerization, and we identify the presence of two intermolecular disulfide bridges between Cys367 and Cys542 and Cys418 and Cys426, respectively. Recently, we reported that the same cysteines contribute to the redox regulation of STAT3 signaling pathway both in vitro and in vivo The presence of these disulfides is here demonstrated to largely contribute to the structure and the stability of U-STAT3 dimer as the dimeric form rapidly dissociates upon reduction in the S-S bonds. In particular, the Cys367-Cys542 disulfide bridge is shown to be critical for U-STAT3 DNA-binding activity. Mutation of the two Cys residues completely abolishes the DNA-binding capability of U-STAT3. Spectroscopic investigations confirm that the noncovalent interactions are sufficient for proper folding and dimer formation, but that the interchain disulfide bonds are crucial to preserve the functional dimer. Finally, we propose a reaction scheme of U-STAT3 dimerization with a first common step followed by stabilization through the formation of interchain disulfide bonds.
authors.affiliation: Department of Neuroscience, Biomedicine and Movement, Biochemistry Section, University of Verona, Verona, Italy.
authors.forename: Elena
authors.initials: E
authors.lastname: Butturini
authors.affiliation: Department of Neuroscience, Biomedicine and Movement, Biochemistry Section, University of Verona, Verona, Italy.
authors.forename: Giovanni
authors.initials: G
authors.lastname: Gotte
authors.affiliation: Department of Neuroscience, Biomedicine and Movement, Biochemistry Section, University of Verona, Verona, Italy.
authors.forename: Daniele
authors.initials: D
authors.lastname: Dell'Orco
authors.affiliation: Department of Neuroscience, Biomedicine and Movement, Biochemistry Section, University of Verona, Verona, Italy.
authors.forename: Giulia
authors.initials: G
authors.lastname: Chiavegato
authors.affiliation: Department of Neuroscience, Biomedicine and Movement, Biochemistry Section, University of Verona, Verona, Italy.
authors.forename: Valerio
authors.initials: V
authors.lastname: Marino
authors.affiliation: CEINGE Biotecnologie Avanzate and Department of Chemical Science, University of Naples 'Federico II', Naples, Italy Istituto Nazionale di Biostrutture e Biosistemi (INBB), Rome, Italy.
authors.forename: Diana
authors.initials: D
authors.lastname: Canetti
authors.affiliation: CEINGE Biotecnologie Avanzate and Department of Chemical Science, University of Naples 'Federico II', Naples, Italy.
authors.forename: Flora
authors.initials: F
authors.lastname: Cozzolino
authors.affiliation: CEINGE Biotecnologie Avanzate and Department of Chemical Science, University of Naples 'Federico II', Naples, Italy.
authors.forename: Maria
authors.initials: M
authors.lastname: Monti
authors.affiliation: CEINGE Biotecnologie Avanzate and Department of Chemical Science, University of Naples 'Federico II', Naples, Italy.
authors.forename: Piero
authors.initials: P
authors.lastname: Pucci
authors.affiliation: Department of Neuroscience, Biomedicine and Movement, Biochemistry Section, University of Verona, Verona, Italy.
authors.forename: Sofia
authors.initials: S
authors.lastname: Mariotto
mesh_terms: D002850:Chromatography, Gel; D019281:Dimerization; D004220:Disulfides / Q000737:chemistry / Q000378:metabolism*; D013058:Mass Spectrometry; D010766:Phosphorylation; D011487:Protein Conformation; D050796:STAT3 Transcription Factor / Q000737:chemistry / Q000378:metabolism*; D020349:Surface Plasmon Resonance; D014443:Tyrosine / Q000378:metabolism
chemical_list: D004220:Disulfides; D050796:STAT3 Transcription Factor; D014443:Tyrosine
keywords: STAT3; dimerization; disulfide bonds