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

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

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

2 Citations (Scopus)

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
Title of host publicationProgress in Biomedical Optics and Imaging - Proceedings of SPIE
Volume8226
DOIs
StatePublished - 2012
EventMultiphoton Microscopy in the Biomedical Sciences XII - San Francisco, CA, United States
Duration: Jan 22 2012Jan 24 2012

Other

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

Fingerprint

Fluorescence Spectrometry
Nuclear Proteins
Fluorescence
Spectroscopy
proteins
Proteins
life (durability)
fluorescence
spectroscopy
interactions
Energy Transfer
Microscopy
Microscopic examination
Heterochromatin
Optical Imaging
Imaging techniques
Gene expression
Energy transfer
Transcription factors
Transcription Factors

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

Cite this

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

Dynamic nuclear protein interactions investigated using fluorescence lifetime and fluorescence fluctuation spectroscopy. / Siegel, Amanda P.; Hays, Nicole M.; Day, Richard.

Progress in Biomedical Optics and Imaging - Proceedings of SPIE. Vol. 8226 2012. 82260B.

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

Siegel, AP, Hays, NM & Day, R 2012, Dynamic nuclear protein interactions investigated using fluorescence lifetime and fluorescence fluctuation spectroscopy. in Progress in Biomedical Optics and Imaging - Proceedings of SPIE. vol. 8226, 82260B, Multiphoton Microscopy in the Biomedical Sciences XII, San Francisco, CA, United States, 1/22/12. https://doi.org/10.1117/12.912883
Siegel AP, Hays NM, Day R. Dynamic nuclear protein interactions investigated using fluorescence lifetime and fluorescence fluctuation spectroscopy. In Progress in Biomedical Optics and Imaging - Proceedings of SPIE. Vol. 8226. 2012. 82260B https://doi.org/10.1117/12.912883
Siegel, Amanda P. ; Hays, Nicole M. ; Day, Richard. / Dynamic nuclear protein interactions investigated using fluorescence lifetime and fluorescence fluctuation spectroscopy. Progress in Biomedical Optics and Imaging - Proceedings of SPIE. Vol. 8226 2012.
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