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

Elizabeth Hinde, Elvis Pandzic, Zhengmin Yang, Ivan H.W. Ng, David A. Jans, Marie A. Bogoyevitch, Enrico Gratton, Katharina Gaus

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19 Citations (Scopus)

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.
Original languageEnglish
Article number11047
Number of pages14
JournalNature Communications
Volume7
DOIs
Publication statusPublished - 24 Mar 2016

Keywords

  • biological sciences
  • cell biology
  • molecular biology

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