Quantifying the dynamics of the oligomeric transcription factor STAT3 by pair correlation of molecular brightness

Elizabeth Hinde (), Elvis Pandžić, Zhengmin Yang, Ivan H. W. Ng, David A. Jans, Marie A. Bogoyevitch, Enrico Gratton and Katharina Gaus ()
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Elizabeth Hinde: EMBL Australia Node in Single Molecule Science, School of Medical Sciences, University of New South Wales
Elvis Pandžić: EMBL Australia Node in Single Molecule Science, School of Medical Sciences, University of New South Wales
Zhengmin Yang: EMBL Australia Node in Single Molecule Science, School of Medical Sciences, University of New South Wales
Ivan H. W. Ng: Program in Emerging Infectious Diseases, Duke-NUS Medical School
David A. Jans: Nuclear Signalling Lab, Faculty of Biomedical and Psychological Sciences, Monash University
Marie A. Bogoyevitch: Cell Signalling Research Laboratories, University of Melbourne
Enrico Gratton: Laboratory for Fluorescence Dynamics, University of California
Katharina Gaus: EMBL Australia Node in Single Molecule Science, School of Medical Sciences, University of New South Wales

Nature Communications, 2016, vol. 7, issue 1, 1-14

Abstract: 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.

Date: 2016
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DOI: 10.1038/ncomms11047

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