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Considering Discrete Protein Pools when Measuring the Dynamics of Nuclear Membrane Proteins

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Imaging Gene Expression

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1042))

Abstract

Measuring dynamics of nuclear proteins is complicated by the fact that many DNA- and chromatin-binding proteins have separate nucleoplasmic and nuclear membrane pools with distinct mobilities. Moreover, when measuring recoveries in FRAP experiments, it is important to be aware that the continuous transport of new protein through the nuclear pore complexes means that fluorescence recovery comes from both dynamic exchange of protein already within the nucleus and newly imported protein. Here we describe fluorescence recovery after photobleaching and photoactivation techniques designed to track nuclear membrane proteins and some methods we have developed that may help to distinguish these various pools. A combination of these approaches with standard FRAP approaches is necessary to understand the true dynamics of nuclear proteins.

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Abbreviations

ER Endoplasmic reticulum FRAP:

Fluorescence recovery after photobleaching

INM Inner nuclear membrane NE:

Nuclear envelope

NET:

Nuclear envelope transmembrane protein

NPC:

Nuclear pore complex ONM Outer nuclear membrane

ROI:

Region of interest

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Acknowledgments

This work was supported by Wellcome Trust Senior Research Fellowship 095209 to E.C.S. and Wellcome Trust Centre for Cell Biology core funding 092076.

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Authors and Affiliations

  1. The Wellcome Trust Centre for Cell Biology, University of Edinburgh, Edinburgh, UK

    Nikolaj Zuleger, David A. Kelly & Eric C. Schirmer

Authors
  1. Nikolaj Zuleger
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  2. David A. Kelly
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  3. Eric C. Schirmer
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Editor information

Editors and Affiliations

  1. Bar-Ilan University Inst. of Nanotech & Advanced Materials, Ramat Gan, Israel

    Yaron Shav-Tal

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Zuleger, N., Kelly, D.A., Schirmer, E.C. (2013). Considering Discrete Protein Pools when Measuring the Dynamics of Nuclear Membrane Proteins. In: Shav-Tal, Y. (eds) Imaging Gene Expression. Methods in Molecular Biology, vol 1042. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-526-2_20

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  • DOI: https://doi.org/10.1007/978-1-62703-526-2_20

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  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-62703-525-5

  • Online ISBN: 978-1-62703-526-2

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