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Binding of calmodulin to Nuf1p is required for karyogamy in Saccharomyces cerevisiae

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Abstract

The role of calmodulin (CaM) during mating in Saccharomyces cerevisiae was examined by using a set of Phe-to-Ala substitutions. We identified ten CaM mutants that exhibited significantly reduced mating efficiencies when crossed to a strain of the opposite mating type harboring the same CaM mutation. Most of the mating-defective CaM mutants were bilateral, i.e., they also exhibited mating defects, albeit minor ones, when crossed to the wild type. When strains carrying different bilateral CaM mutations were mated, the mating efficiencies recovered dramatically. We termed this phenomenon "intragenic mating complementation", and classified the mating-defective CaM mutations into two intragenic mating complementation groups. Two mutant alleles belonging to different groups showed minor defects in cell adhesion and cell fusion, but exhibited severe defects in karyogamy. CaM is known to bind to the essential spindle pole body component Nuf1p. This binding appears to be important for karyogamy because the nuf1 C911R mutation, which impairs CaM-Nuf1p binding, resulted in a severe defect in karyogamy. Indeed, the two mating-defective CaM mutations were found to compromise formation of the CaM/Nuf1p complex, and the mating defects of these two CaM mutants were suppressible by a dominant, CaM-independent, mutation in NUF1. Taken together, these results suggest that loss of CaM binding to Nuf1p causes a defect in karyogamy, thereby inhibiting productive mating.

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Acknowledgements

We would like to thank Hidenori Kimura for encouragement throughout this study, Rino Matsunaga for critically reading the manuscript, and members of the Laboratory of Signal Transduction for helpful discussion. This work was supported by grants from the Ministry of Education, Science and Sports and Culture of Japan to Y. Ohya, and from the Japan Society for the Promotion of Science for Young Scientists to H. Okano

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Authors and Affiliations

  1. Bio-Mimetic Control Research Center, The Institute of Physical and Chemical Research RIKEN, Anagahora, 463-0003, Nagoya, Aichi, Japan

    H. Okano

  2. Department of Integrated Biosciences, Graduate School of Frontier Sciences, University of Tokyo, Bldg. FSB-101, 5-1-5 Kashiwanoha, 277-8562, Kashiwa, Chiba, Japan

    Y. Ohya

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  1. H. Okano
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  2. Y. Ohya
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Correspondence to Y. Ohya.

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Communicated by C. P. Hollenberg

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Okano, H., Ohya, Y. Binding of calmodulin to Nuf1p is required for karyogamy in Saccharomyces cerevisiae . Mol Gen Genomics 269, 649–657 (2003). https://doi.org/10.1007/s00438-003-0853-z

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