Dynamic nuclear protein interactions investigated using fluorescence lifetime and fluorescence fluctuation spectroscopy

Amanda P. Siegel, Nicole M. Hays, Richard N. Day

Research output: Chapter in Book/Report/Conference proceedingConference contribution

2 Scopus citations

Abstract

The discovery and engineering of novel fluorescent proteins (FPs) from diverse organisms is yielding fluorophores with exceptional characteristics for live-cell imaging. In particular, the development of FPs for Förster resonance energy transfer (FRET) microscopy and fluorescence fluctuation spectroscopy (FFS) provide important tools for monitoring dynamic protein interactions inside living cells. Fluorescence lifetime imaging microscopy (FLIM) quantitatively maps changes in the spatial distribution of donor FP lifetimes that result from FRET with acceptor FPs. FFS probes dynamic protein associations through its capacity to monitor localized protein diffusion. Here, we use FRET-FLIM combined with FFS in living cells to investigate changes in protein mobility due to protein-protein interactions involving transcription factors and chromatin modifying proteins that function in anterior pituitary gene regulation. The heterochromatin protein 1 alpha (HP1α) plays a key role in the establishment and maintenance of heterochromatin through its interactions with histone methyltransferases. Recent studies, however, also highlight the importance of HP1α as a positive regulator of active transcription in euchromatin. Intriguingly, we observed that the transcription factor CCAAT/enhancer-binding protein alpha (C/EBPα) interacts with HP1α in regions of pericentromeric heterochromatin in mouse pituitary cells. These observations prompted us to investigate the relationship between HP1α dynamic interactions in pituitary specific gene regulation.

Original languageEnglish (US)
Title of host publicationMultiphoton Microscopy in the Biomedical Sciences XII
DOIs
StatePublished - Apr 16 2012
EventMultiphoton Microscopy in the Biomedical Sciences XII - San Francisco, CA, United States
Duration: Jan 22 2012Jan 24 2012

Publication series

NameProgress in Biomedical Optics and Imaging - Proceedings of SPIE
Volume8226
ISSN (Print)1605-7422

Other

OtherMultiphoton Microscopy in the Biomedical Sciences XII
CountryUnited States
CitySan Francisco, CA
Period1/22/121/24/12

Keywords

  • fluorescence fluctuation spectroscopy (FFS)
  • fluorescence lifetime imaging microscopy (FLIM)
  • fluorescence resonance energy transfer (FRET) microscopy
  • fluorescent proteins (FPs)
  • protein interactions

ASJC Scopus subject areas

  • Atomic and Molecular Physics, and Optics
  • Electronic, Optical and Magnetic Materials
  • Biomaterials
  • Radiology Nuclear Medicine and imaging

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  • Cite this

    Siegel, A. P., Hays, N. M., & Day, R. N. (2012). Dynamic nuclear protein interactions investigated using fluorescence lifetime and fluorescence fluctuation spectroscopy. In Multiphoton Microscopy in the Biomedical Sciences XII [82260B] (Progress in Biomedical Optics and Imaging - Proceedings of SPIE; Vol. 8226). https://doi.org/10.1117/12.912883