Scientific Reports (May 2021)

Live cell dynamics of the NF-Y transcription factor

  • David G. Priest,
  • Andrea Bernardini,
  • Jieqiong Lou,
  • Roberto Mantovani,
  • Elizabeth Hinde

DOI
https://doi.org/10.1038/s41598-021-90081-1
Journal volume & issue
Vol. 11, no. 1
pp. 1 – 10

Abstract

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Abstract Transcription factors (TFs) are core players in the control of gene expression, evolutionarily selected to recognise a subset of specific DNA sequences and nucleate the recruitment of the transcriptional machinery. How TFs assemble and move in the nucleus to locate and bind their DNA targets and cause a transcriptional response, remains mostly unclear. NF-Y is a highly conserved, heterotrimeric TF with important roles in both housekeeping and lineage-specific gene expression, functioning as a promoter organiser. Despite a large number of biochemical, structural and genomic studies of NF-Y, there is a lack of experiments in single living cells; therefore, basic assumptions of NF-Y biology remain unproven in vivo. Here we employ a series of dynamic fluorescence microscopy methods (FLIM-FRET, NB, RICS and FRAP) to study NF-Y dynamics and complex formation in live cells. Specifically, we provide quantitative measurement of NF-Y subunit association and diffusion kinetics in the nucleus that collectively suggest NF-Y to move and bind chromatin as a trimeric complex in vivo.