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Functional Analysis of the Yeast LINC Complex Using Fluctuation Spectroscopy and Super-Resolution Imaging

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The LINC Complex

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1840))

Abstract

The Saccharomyces cerevisiae and Schizosaccharomyces pombe genomes encode a single SUN domain-containing protein, Mps3 and Sad1, respectively. Both localize to the yeast centrosome (known as the spindle pole body, SPB) and are essential for bipolar spindle formation. In addition, Mps3 and Sad1 play roles in chromosome organization in both mitotic and meiotic cells that are independent of their SPB function. To dissect the function of Mps3 at the nuclear envelope (NE) and SPB, we employed cell imaging methods such as scanning fluorescence cross-correlation spectroscopy (SFCCS) and single particle averaging with structured illumination microscopy (SPA-SIM) to determine the strength, nature, and location of protein-protein interactions in vivo. We describe how these same techniques can also be used in fission yeast to analyze Sad1, providing evidence of their applicability to other NE proteins and systems.

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Acknowledgments

We are grateful to members of the Microscopy Center and the Jaspersen lab for stimulating discussions and to Briana Holt, Jennifer Gardner, Christine Smoyer, Ann Cavanaugh, Andrew Bestul, and Jeff Lange for their comments on the manuscript.

Author information

Authors and Affiliations

  1. Stowers Institute for Medical Research, Kansas City, MO, USA

    Jay R. Unruh, Brian D. Slaughter & Sue L. Jaspersen

  2. Department of Molecular and Integrative Physiology, University of Kansas Medical Center, Kansas City, KS, USA

    Sue L. Jaspersen

Authors
  1. Jay R. Unruh
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  2. Brian D. Slaughter
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  3. Sue L. Jaspersen
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Corresponding author

Correspondence to Sue L. Jaspersen .

Editor information

Editors and Affiliations

  1. Department of Pathology and Cell Biology, Columbia University, New York, New York, USA

    Gregg G. Gundersen

  2. Department of Medicine and Department of Pathology and Cell Biology, Columbia University, New York, New York, USA

    Howard J. Worman

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Unruh, J.R., Slaughter, B.D., Jaspersen, S.L. (2018). Functional Analysis of the Yeast LINC Complex Using Fluctuation Spectroscopy and Super-Resolution Imaging. In: Gundersen, G., Worman, H. (eds) The LINC Complex. Methods in Molecular Biology, vol 1840. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-8691-0_12

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  • DOI: https://doi.org/10.1007/978-1-4939-8691-0_12

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-8690-3

  • Online ISBN: 978-1-4939-8691-0

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