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Coupling of cortical dynein and Gα proteins mediates spindle positioning in Caenorhabditis elegans

Nature Cell Biology volume 9, pages 1294–1302 (2007)Cite this article

Abstract

Despite being essential for spatial cell division control, the mechanisms governing spindle positioning remain incompletely understood. In the Caenorhabditis elegans one-cell stage embryo, the spindle becomes asymmetrically positioned during anaphase through the action of as-yet unidentified cortical force generators that pull on astral microtubules and that depend on two Gα proteins and associated proteins1,2. We performed spindle-severing experiments following temporally restricted gene inactivation and drug exposure, and established that microtubule dynamics and dynein are both required for generating efficient pulling forces. We found that the Gα-associated proteins GPR-1/2 and LIN-5 interact in vivo with LIS-1, a component of the dynein complex. Moreover, we discovered that the LIN-5, GPR-1/2 and the Gα proteins promote the presence of the dynein complex at the cell cortex. Our findings suggest a mechanism by which the Gα proteins enable GPR-1/2 and LIN-5 recruitment to the cortex, thus ensuring the presence of cortical dynein. Together with microtubule dynamics, this allows pulling forces to be exerted and proper cell division to be achieved.

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Figure 1: Proper microtubule dynamics are essential for pulling forces.
Figure 2: Dynein and LIN-5 contribute to pulling forces and interact with each other in vivo.
Figure 3: Cortical DHC-1 is dependent on LIN-5, GPR-1/2 and Gα.
Figure 4: Cortical LIS-1 is dependent on LIN-5, GPR-1/2 and Gα (a–h) Wild-type, lin-5(RNAi), gpr-1/2(RNAi) and Gα(RNAi) one-cell and two-cell stage embryos, as indicated, stained with antibodies against LIS-1 (red), α-tubulin (green) and counterstained with Hoechst to view DNA (blue).
Figure 5: Cortical dynein couples force generation with Gα proteins.

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Acknowledgements

We are grateful to Daniel Constam, Virginie Hachet and Kalyani Thyagarajan for critical reading of the manuscript, Virginie Hachet for affinity purifying GPR-1/2 antibodies, John Blanc and Hugues Baumgartner for building the temperature-shift device, Claude Bonnard and Natsuko Imaizumi for help with image processing. Mutant and transgenic strains were kindly provided by Bruce Bowerman, Craig Hunter, Anthony Hyman and Sander van den Heuvel. Supported by grant 3100A0-102087 from the Swiss National Science Foundation.

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  1. Swiss Institute for Experimental Cancer Research (ISREC), Swiss Federal Institute of Technology (EPFL), School of Life Sciences, Lausanne, CH-1066, Switzerland

    Tu Nguyen-Ngoc, Katayoun Afshar & Pierre Gönczy

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  1. Tu Nguyen-Ngoc
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Correspondence to Pierre Gönczy.

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Nguyen-Ngoc, T., Afshar, K. & Gönczy, P. Coupling of cortical dynein and Gα proteins mediates spindle positioning in Caenorhabditis elegans. Nat Cell Biol 9, 1294–1302 (2007). https://doi.org/10.1038/ncb1649

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