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Deletion of RNQ1 gene reveals novel functional relationship between divergently transcribed Bik1p/CLIP-170 and Sfi1p in spindle pole body separation

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Abstract

Spindle pole body (SPB; the microtubule organizing center in yeast) duplication is essential to form a bipolar spindle. The duplicated SPBs must then separate and migrate to opposite sides of the nucleus. We identified a novel functional relationship in SPB separation between the microtubule stabilizing protein Bik1p/CLIP-170 and the SPB half-bridge protein Sfi1p. A genetic interaction between BIK1 and SFI1 was discovered in a synthetic lethal screen using a strain deficient in the prion protein gene RNQ1. RNQ1 deletion reduced expression from the divergently transcribed BIK1, allowing us to identify genetic interactors with bik1. The sfi1-1 bik1 synthetic lethality was suppressed by over-expression of CIK1, KAR1, and PPH21. Genetic analysis indicated that the sfi1-1 bik1 synthetic lethality was unlikely related to the function of Bik1p in the dynein pathway or to defects in spindle position. Furthermore, a sfi1-1 Δkip2 mutant was viable, suggesting that the Bik1p pool at the cytoplasmic microtubule plus-ends may not be required in sfi1-1. Microscopic examination indicated the sfi1-1 mutant was delayed in SPB duplication, SPB separation, or spindle elongation and the sfi-1 Δbik1 double mutant arrested with duplicated but unseparated SPBs. These results suggest that Bik1p has a previously uncharacterized function in the separation of duplicated SPBs.

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Acknowledgments

We gratefully acknowledge Marissa Matsumoto for initiating the synthetic lethal screen, Sarah Broekelmann for technical support, Larry Schriefer for assistance with DNA sequencing, and Howard Wynder for assistance with EM. We thank Anthony Bretscher, John Cooper, John Kilmartin, Martin Kupiec, Susan Lindquist, Rita Miller, and Mark Rose for sharing plasmids, strains, antibodies, libraries, and equipment. We are grateful to John Cooper, Jun Li, Scott Nelson, Mark Winey, and members of the True lab for helpful discussions. We thank Rachel Boutenott, Jun Li, and Scott Nelson for providing critical comments on the manuscript. This work was supported by funds from National Institutes of Health Grant AG024560, the Edward Mallinckrodt Jr. Foundation, and the Ellison Medical Foundation (awarded to H.L.T.). L.A.S. was supported by a fellowship from the Washington University Infectious Diseases Scholars Program (National Institutes of Health Ruth L. Kirschstein National Research Service Award Training Grant T32 A107172-25) and a USDA National Research Initiative Postdoctoral AREA Award (2005-35201-15383).

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  1. Department of Cell Biology and Physiology, Washington University School of Medicine, 660 S Euclid Ave, Campus Box 8228, St Louis, MO, 63110, USA

    Lisa A. Strawn & Heather L. True

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  1. Lisa A. Strawn
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Correspondence to Heather L. True.

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Communicated by M. Kupiec.

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Strawn, L.A., True, H.L. Deletion of RNQ1 gene reveals novel functional relationship between divergently transcribed Bik1p/CLIP-170 and Sfi1p in spindle pole body separation. Curr Genet 50, 347–366 (2006). https://doi.org/10.1007/s00294-006-0098-6

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