Live cell dynamics of the NF-Y transcription factor.

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

Scientific reports 2021 May 26

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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.

Citation

David G Priest, Andrea Bernardini, Jieqiong Lou, Roberto Mantovani, Elizabeth Hinde. Live cell dynamics of the NF-Y transcription factor. Scientific reports. 2021 May 26;11(1):10992