PMID: 8601600Apr 1, 1996Paper

Role of calmodulin and Spc110p interaction in the proper assembly of spindle pole body compenents

The Journal of Cell Biology
H A SundbergT N Davis

Abstract

Previously we demonstrated that calmodulin binds to the carboxy terminus of Spc110p, an essential component of the Saccharomyces cerevisiae spindle pole body (SPB), and that this interaction is required for chromosome segregation. Immunoelectron microscopy presented here shows that calmodulin and thus the carboxy terminus of Spc110p localize to the central plaque. We created temperature-sensitive SPC110 mutations by combining PCR mutagenesis with a plasmid shuffle strategy. The temperature-sensitive allele spc110-220 differs from wild type at two sites. The cysteine 911 to arginine mutation resides in the calmodulin-binding site and alone confers a temperature-sensitive phenotype. Calmodulin overproduction suppresses the temperature sensitivity of spc110-220. Furthermore, calmodulin levels at the SPB decrease in the mutant cells at the restrictive temperature. Thus, calmodulin binding to Spc110-220p is defective at the nonpermissive temperature. Synchronized mutant cells incubated at the nonpermissive temperature arrest as large budded cells with a G2 content of DNA and suffer considerable lethality. Immunofluorescent staining demonstrates failure of nuclear DNA segregation and breakage of many spindles. Electron microscopy rev...Continue Reading

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Citations

Feb 1, 1997·The Journal of Experimental Zoology·S G Sobel
May 14, 1999·Current Opinion in Structural Biology·C Wiese, Y Zheng
Jun 25, 1998·Current Biology : CB·K TörökM Whitaker
Aug 1, 1997·Zygote : the Biology of Gametes and Early Embryos·M A HenryM M Fahy
May 15, 2010·The Journal of Biological Chemistry·Zhe WangRobert Z Qi
Apr 16, 2002·Molecular Biology of the Cell·Dani B N VinhTrisha N Davis
Nov 1, 1996·Molecular Biology of the Cell·R A SiaD J Lew
Aug 20, 2003·Molecular Biology of the Cell·Tennessee J YoderTrisha N Davis
Oct 21, 2003·Molecular Biology of the Cell·Shin-ichi Kawaguchi, Yixian Zheng
Nov 5, 2004·Molecular Biology of the Cell·Tennessee J YoderTrisha N Davis
Jun 9, 2006·Molecular Biology of the Cell·William Y TsangBrian David Dynlacht
Dec 6, 2008·Molecular Biology of the Cell·Michelle M ShimogawaTrisha N Davis
Dec 21, 2007·Biology of Reproduction·Lu SunKhaled Machaca
Oct 12, 2004·Annual Review of Cell and Developmental Biology·Sue L Jaspersen, Mark Winey
Jan 1, 1996·Annual Review of Genetics·M A Hoyt, J R Geiser
Nov 26, 2010·PloS One·Kristen B GreenlandTrisha N Davis
Apr 12, 2012·Genetics·Mark Winey, Kerry Bloom
May 26, 1999·Proceedings of the National Academy of Sciences of the United States of America·S ElliottE Schiebel
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Dec 3, 2009·The Journal of Cell Biology·Benedicte Delaval, Stephen J Doxsey
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Sep 23, 1998·Experimental Cell Research·C Gueth-HallonetM Osborn
Nov 18, 1997·Cell·T Stearns, M Winey
May 27, 2016·Molecular Biology of the Cell·Andrew S LyonDavid A Agard

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