The molecular architecture of the yeast spindle pole body core determined by Bayesian integrative modeling

Viswanath, Shruthi; Bonomi, Massimiliano; Kim, Seung Joong; Klenchin, Vadim A.; Taylor, Keenan C.; Yabut, King C.; Umbreit, Neil T.; Van Epps, Heather A.; Meehl, Janet; Jones, Michele H.; Russel, Daniel; Winey, Mark; Rayment, Ivan; Davis, Trisha N.; Sali, Andrej; Muller, Eric G.; Bloom, Kerry S.; Velazquez-Muriela, Javier A.

doi:10.1091/mbc.e17-06-0397

Title: The molecular architecture of the yeast spindle pole body core determined by Bayesian integrative modeling

Journal Article · 15 August 2017 · Molecular Biology of the Cell

DOI: https://doi.org/10.1091/mbc.e17-06-0397 · OSTI ID:1625244

Viswanath, Shruthi ^[1]; Bonomi, Massimiliano ^[2]; Kim, Seung Joong ^[3]; Klenchin, Vadim A. ^[4]; Taylor, Keenan C. ^[5]; Yabut, King C. ^[6]; Umbreit, Neil T. ^[7]; Van Epps, Heather A. ^[7]; Meehl, Janet ^[8]; Jones, Michele H. ^[8]; Russel, Daniel ^[9]; Winey, Mark ^[8]; Rayment, Ivan ^[5]; Davis, Trisha N. ^[7]; Sali, Andrej ^[3]; Muller, Eric G. ^[7]; Bloom, Kerry S. ^[10]; Velazquez-Muriela, Javier A. ^[3]

Univ. of California, San Francisco, CA (United States). Department of Bioengineering and Therapeutic Sciences, Department of Pharmaceutical Chemistry, California Institute for Quantitative Biosciences; DOE/OSTI
Univ. of California, San Francisco, CA (United States). Department of Bioengineering and Therapeutic Sciences, Department of Pharmaceutical Chemistry, California Institute for Quantitative Biosciences; Univ. of Cambridge (United Kingdom). Department of Chemistry
Univ. of California, San Francisco, CA (United States). Department of Bioengineering and Therapeutic Sciences, Department of Pharmaceutical Chemistry, California Institute for Quantitative Biosciences
Univ. of Wisconsin-Madison, Madison, WI (United States). Department of Biochemistry
Univ. of Wisconsin-Madison, Madison, WI (United States). Department of Biochemistry
Univ. of Washington, Seattle, WA (United States). Dept. of Biochemistry
Univ. of Washington, Seattle, WA (United States). Dept. of Biochemistry
Univ. of Colorado-Boulder, Boulder, CO (United States). Dept. of Molecular, Cellular and Developmental Biology
Univ. of California, San Francisco, CA (United States). Department of Bioengineering and Therapeutic Sciences, Department of Pharmaceutical Chemistry, California Institute for Quantitative Biosciences
Univ. of North Carolina, Chapel Hill, NC (United States)

Microtubule-organizing centers (MTOCs) form, anchor, and stabilize the polarized network of microtubules in a cell. The central MTOC is the centrosome that duplicates during the cell cycle and assembles a bipolar spindle during mitosis to capture and segregate sister chromatids. Yet, despite their importance in cell biology, the physical structure of MTOCs is poorly understood. Here we determine the molecular architecture of the core of the yeast spindle pole body (SPB) by Bayesian integrative structure modeling based on in vivo fluorescence resonance energy transfer (FRET), small-angle x-ray scattering (SAXS), x-ray crystallography, electron microscopy, and two-hybrid analysis. The model is validated by several methods that include a genetic analysis of the conserved PACT domain that recruits Spc110, a protein related to pericentrin, to the SPB. The model suggests that calmodulin can act as a protein cross-linker and Spc29 is an extended, flexible protein. The model led to the identification of a single, essential heptad in the coiled-coil of Spc110 and a minimal PACT domain. It also led to a proposed pathway for the integration of Spc110 into the SPB.

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Research Organization:: SLAC National Accelerator Laboratory, Menlo Park, CA (United States). Stanford Synchrotron Radiation Lightsource (SSRL)

Sponsoring Organization:: USDOE Office of Science (SC), Basic Energy Sciences (BES); Office of Science (SC), Biological and Environmental Research (BER)

Grant/Contract Number:: AC02-76SF00515

OSTI ID:: 1625244

Journal Information:: Molecular Biology of the Cell, Journal Name: Molecular Biology of the Cell Journal Issue: 23 Vol. 28; ISSN 1059-1524

Publisher:: American Society for Cell BiologyCopyright Statement

Country of Publication:: United States

Language:: English

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