Letter | Published:

Cortical forces and CDC-42 control clustering of PAR proteins for Caenorhabditis elegans embryonic polarization

Nature Cell Biology volume 19, pages 988995 (2017) | Download Citation

Abstract

Cell polarization enables zygotes to acquire spatial asymmetry, which in turn patterns cellular and tissue axes during development. Local modification in the actomyosin cytoskeleton mediates spatial segregation of partitioning-defective (PAR) proteins at the cortex1,2,3, but how mechanical changes in the cytoskeleton are transmitted to PAR proteins remains elusive. Here we uncover a role of actomyosin contractility in the remodelling of PAR proteins through cortical clustering. During embryonic polarization in Caenorhabditis elegans, actomyosin contractility and the resultant cortical tension stimulate clustering of PAR-3 at the cortex. Clustering of atypical protein kinase C (aPKC) is supported by PAR-3 clusters and is antagonized by activation of CDC-42. Cortical clustering is associated with retardation of PAR protein exchange at the cortex and with effective entrainment of advective cortical flows. Our findings delineate how cytoskeleton contractility couples the cortical clustering and long-range displacement of PAR proteins during polarization. The principles described here would apply to other pattern formation processes that rely on local modification of cortical actomyosin and PAR proteins.

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Acknowledgements

This study was supported by the Singapore National Research Foundation (NRF_NRFF2012-08 (F.M.)) and the Strategic Japan-Singapore Cooperative Research Program by the Japan Science and Technology Agency and the Singapore Agency for Science, Technology, and Research (1514324022 (F.M.)). We are grateful to B. Goldstein and D. Dickinson (The University of North Carolina at Chapel Hill), K. Kemphues (Cornell University), J. C. F. Li, F. Margadant, H. T. Ong and A. Bershadsky (Mechanobiology Institute, Singapore), G. Seydoux (Johns Hopkins University), S. Mathew (Temasek Life-sciences Laboratory, Singapore), and the Caenorhabditis Genetic Center for strains, reagents and expertise. We thank B. Goldstein, D. Dickinson and N. Goehring (The Francis Crick Institute) for sharing their results with us before publication. We also thank A. Wong (Mechanobiology Institute, Singapore) and members in the Motegi laboratory for helpful comments on the manuscript.

Author information

Author notes

    • Tricia Yu Feng Low
    • , Yukako Nishimura
    •  & Laurent Gole

    These authors contributed equally to this work.

Affiliations

  1. Temasek Life-sciences Laboratory, 1 Research Link, National University of Singapore, Singapore 117604, Singapore

    • Shyi-Chyi Wang
    • , Tricia Yu Feng Low
    •  & Fumio Motegi
  2. Mechanobiology Institute, National University of Singapore, T-Lab, 5A Engineering Drive 1, Singapore 117411, Singapore

    • Yukako Nishimura
    •  & Fumio Motegi
  3. Institute for Molecular and Cell Biology, Agency for Science Technology and Research, 61 Biopolis Drive, Singapore 138673, Singapore

    • Laurent Gole
    •  & Weimiao Yu
  4. Department of Biological Sciences, National University of Singapore, 14 Science Drive 4, Singapore 117543, Singapore

    • Fumio Motegi

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Contributions

The experimental design and presented ideas were developed together by all authors. F.M. guided the study and wrote the manuscript with input from all authors. S.-C.W. performed experiments in Figs 1, 3 and 4a, b and Supplementary Figs 1, 3 and 4a. T.Y.F.L. performed experiments in Figs 2 and 4c–g and Supplementary Figs 2 and 4b. Y.N. performed experiments in Fig. 2i–k and Supplementary Fig. 2c, d. L.G. and W.Y. developed program codes for PIV analysis and cortical cluster quantification, and analysed videos shown in Fig. 4c–g and Supplementary Fig. 4b.

Competing interests

The authors declare no competing financial interests.

Corresponding authors

Correspondence to Weimiao Yu or Fumio Motegi.

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Videos

  1. 1.

    Cortical PAR-3::GFP with mCherry::PAR-6 in a one-cell stage wild-type zygote.

    Scale bar, 5 μm. Related to Fig. 1a, c, d, g and Supplementary Fig. 1b, c, f.

  2. 2.

    Cortical PKC-3::GFP with mCherry::PAR-6 in a one-cell stage wild-type zygote.

    Scale bar, 5 μm. Related to Fig. 1a, e, f, g and Supplementary Fig. 1c, f.

  3. 3.

    Cortical GFP::CDC-42 with mCherry::PAR-6 in a one-cell stage wild-type zygote.

    Scale bar, 5 μm. Related to Fig. 1a, g.

  4. 4.

    Cortical PAR-3::GFP, PKC-3::GFP and GFP::CDC-42 with mCherry::PHPLC in one-cell stage wild-type zygotes.

    Images show the anteromedial cortex and are aligned to disassembly of cortical clusters and cortical foci. Scale bar, 5 μm. Related to Fig. 1b, g.i and Supplementary Fig. 1d, e.

  5. 5.

    Cortical PKC-3::GFP and NMY-2::Kate during cytokinesis phase in a plk-1(or683) zygote.

    Images show the medial cortex. Scale bar, 5 μm. Related to Fig. 2b, c.

  6. 6.

    Cortical PAR-3::GFP in nmy-2(ne3409) zygotes treated with or without hypotonic buffer.

    Images show the anteromedial cortex. Scale bar, 5 μm. Related to Fig. 2d, e, f and Supplementary Fig. 2b, c.

  7. 7.

    Cortical PAR-3::GFP during establishment phases in control and mCherry::CDC-42Q61L expressing zygotes.

    Images show the anteromedial cortex and are aligned to disassembly of cortical clusters. Scale bar, 5 μm. Related to Fig. 3b, c and Supplementary Fig. 3a.

  8. 8.

    Cortical PKC-3::GFP during establishment phases in control and mCherry::CDC-42Q61L expressing zygotes.

    Images show the anteromedial cortex and are aligned to disassembly of cortical clusters. Scale bar, 5 μm. Related to Fig. 3b, d and Supplementary Fig. 3a.

  9. 9.

    Cortical PAR-3::GFP during establishment phases in control, cdc-42(RNAi), and cgef-1(RNAi) zygotes.

    Images show the anteromedial cortex and are aligned to disassembly of cortical clusters. Scale bar, 5 μm. Related to Fig. 3b, e and Supplementary Fig. 3a.

  10. 10.

    Cortical PKC-3::GFP during establishment phases in control, cdc-42(RNAi), and cgef-1(RNAi) zygotes.

    Images show the anteromedial cortex and are aligned to disassembly of cortical clusters. Scale bar, 5 μm. Related to Fig. 3b, f and Supplementary Fig. 3a.

  11. 11.

    Cortical PAR-3::GFP and mCherry::PHPLC during establishment phases in control and cdc-42(RNAi) zygotes.

    Images show the anteromedial cortex and are aligned to disassembly of cortical clusters. Inverted PAR-3::GFP images are shown above in parallel. Scale bar, 5 μm. Related to Fig. 1i and Fig. 3g.

  12. 12.

    Cortical PKC-3::GFP and mCherry::PHPLC during establishment phases in control and cdc-42(RNAi) zygotes.

    Images show the anteromedial cortex and are aligned to disassembly of cortical clusters. Inverted PKC-3::GFP images are shown above in parallel. Scale bar, 5 μm. Related to Fig. 1i and Fig. 3i.

  13. 13.

    Representative FRAP images of PAR-3::GFP and PKC-3::GFP during the early establishment phase in wild-type zygotes.

    Images show the anteromedial cortex, where a 5 μm diameter circular area was photo-bleached, and are aligned to the onset of photo-bleaching. Scale bar, 5 μm. Related to Fig. 4a, b and Supplementary Fig. 4a.

  14. 14.

    PIV analysis of cortical PAR-3::GFP and NMY-2::Kate during establishment phases in wild-type zygote.

    Cortical PAR-3::GFP and NMY-2::Kate, segmented clusters of PAR-3::GFP, PIV vector maps, PIV magnitudes for PAR-3::GFP and NMY-2::Kate, as well as cross-correlation of two PIV vectors orientations are shown synchronically. Scale bar, 5 μm. Related to Fig. 4c–g and Supplementary Fig. 4b.

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DOI

https://doi.org/10.1038/ncb3577