title: Quantifying the dynamics of the oligomeric transcription factor STAT3 by pair correlation of molecular brightness.
abstract: Oligomerization of transcription factors controls their translocation into the nucleus and DNA-binding activity. Here we present a fluorescence microscopy analysis termed pCOMB (pair correlation of molecular brightness) that tracks the mobility of different oligomeric species within live cell nuclear architecture. pCOMB amplifies the signal from the brightest species present and filters the dynamics of the extracted oligomeric population based on arrival time between two locations. We use this method to demonstrate a dependence of signal transducer and activator of transcription 3 (STAT3) mobility on oligomeric state. We find that on entering the nucleus STAT3 dimers must first bind DNA to form STAT3 tetramers, which are also DNA-bound but exhibit a different mobility signature. Examining the dimer-to-tetramer transition by a cross-pair correlation analysis (cpCOMB) reveals that chromatin accessibility modulates STAT3 tetramer formation. Thus, the pCOMB approach is suitable for mapping the impact oligomerization on transcription factor dynamics.
authors.affiliation: EMBL Australia Node in Single Molecule Science, School of Medical Sciences, University of New South Wales, Sydney, New South Wales 2052, Australia.|ARC Centre of Excellence in Advanced Molecular Imaging, University of New South Wales, Sydney, New South Wales 2052, Australia.
authors.forename: Elizabeth
authors.initials: E
authors.lastname: Hinde
authors.affiliation: EMBL Australia Node in Single Molecule Science, School of Medical Sciences, University of New South Wales, Sydney, New South Wales 2052, Australia.|ARC Centre of Excellence in Advanced Molecular Imaging, University of New South Wales, Sydney, New South Wales 2052, Australia.
authors.forename: Elvis
authors.initials: E
authors.lastname: Pandžić
authors.affiliation: EMBL Australia Node in Single Molecule Science, School of Medical Sciences, University of New South Wales, Sydney, New South Wales 2052, Australia.|ARC Centre of Excellence in Advanced Molecular Imaging, University of New South Wales, Sydney, New South Wales 2052, Australia.
authors.forename: Zhengmin
authors.initials: Z
authors.lastname: Yang
authors.affiliation: Program in Emerging Infectious Diseases, Duke-NUS Medical School, Singapore 169587, Singapore.|Nuclear Signalling Lab, Department of Biochemistry and Molecular Biology, Faculty of Biomedical and Psychological Sciences, Monash University, Melbourne, Victoria 3800, Australia.
authors.forename: Ivan H W
authors.initials: IH
authors.lastname: Ng
authors.affiliation: Nuclear Signalling Lab, Department of Biochemistry and Molecular Biology, Faculty of Biomedical and Psychological Sciences, Monash University, Melbourne, Victoria 3800, Australia.
authors.forename: David A
authors.initials: DA
authors.lastname: Jans
authors.affiliation: Cell Signalling Research Laboratories, Department of Biochemistry and Molecular Biology, University of Melbourne, Melbourne, Victoria 3010, Australia.
authors.forename: Marie A
authors.initials: MA
authors.lastname: Bogoyevitch
authors.affiliation: Laboratory for Fluorescence Dynamics, Department of Biomedical Engineering, University of California, Irvine, California 92697-2715, USA.
authors.forename: Enrico
authors.initials: E
authors.lastname: Gratton
authors.affiliation: EMBL Australia Node in Single Molecule Science, School of Medical Sciences, University of New South Wales, Sydney, New South Wales 2052, Australia.|ARC Centre of Excellence in Advanced Molecular Imaging, University of New South Wales, Sydney, New South Wales 2052, Australia.
authors.forename: Katharina
authors.initials: K
authors.lastname: Gaus
mesh_terms: D002138:Calibration; D002470:Cell Survival / Q000187:drug effects; D002843:Chromatin / Q000378:metabolism; D016207:Cytokines / Q000494:pharmacology; D004247:DNA / Q000378:metabolism; D006367:HeLa Cells; D006801:Humans; D008856:Microscopy, Fluorescence / Q000379:methods*; D011485:Protein Binding / Q000187:drug effects; D055503:Protein Multimerization / Q000187:drug effects; D021381:Protein Transport / Q000187:drug effects; D050796:STAT3 Transcription Factor / Q000378:metabolism*
chemical_list: D002843:Chromatin; D016207:Cytokines; D050796:STAT3 Transcription Factor; D004247:DNA