Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Letter
  • Published:

The mitotic checkpoint complex binds a second CDC20 to inhibit active APC/C

Subjects

Abstract

The spindle assembly checkpoint (SAC) maintains genomic stability by delaying chromosome segregation until the last chromosome has attached to the mitotic spindle. The SAC prevents the anaphase promoting complex/cyclosome (APC/C) ubiquitin ligase from recognizing cyclin B and securin by catalysing the incorporation of the APC/C co-activator, CDC20, into a complex called the mitotic checkpoint complex (MCC). The SAC works through unattached kinetochores generating a diffusible ‘wait anaphase’ signal1,2 that inhibits the APC/C in the cytoplasm, but the nature of this signal remains a key unsolved problem. Moreover, the SAC and the APC/C are highly responsive to each other: the APC/C quickly targets cyclin B and securin once all the chromosomes attach in metaphase, but is rapidly inhibited should kinetochore attachment be perturbed3,4. How this is achieved is also unknown. Here, we show that the MCC can inhibit a second CDC20 that has already bound and activated the APC/C. We show how the MCC inhibits active APC/C and that this is essential for the SAC. Moreover, this mechanism can prevent anaphase in the absence of kinetochore signalling. Thus, we propose that the diffusible ‘wait anaphase’ signal could be the MCC itself, and explain how reactivating the SAC can rapidly inhibit active APC/C.

This is a preview of subscription content, access via your institution

Access options

Rent or buy this article

Prices vary by article type

from$1.95

to$39.95

Prices may be subject to local taxes which are calculated during checkout

Figure 1: Core MCC can inhibit APC/CCDC20.
Figure 2: The MCC binds to CDC20 through substrate recognition domains.
Figure 3: MCC binds to the second CDC20 through the D-box of BUBR1 and this is required for the SAC.
Figure 4: A stabilized MCC delays anaphase by inhibiting endogenous APC/CCDC20.

Similar content being viewed by others

References

  1. Rieder, C. L., Cole, R. W., Khodjakov, A. & Sluder, G. The checkpoint delaying anaphase in response to chromosome monoorientation is mediated by an inhibitory signal produced by unattached kinetochores. J. Cell Biol. 130, 941–948 (1995)

    Article  CAS  Google Scholar 

  2. Rieder, C. L. et al. Mitosis in vertebrate somatic cells with two spindles: implications for the metaphase/anaphase transition checkpoint and cleavage. Proc. Natl Acad. Sci. USA 94, 5107–5112 (1997)

    Article  ADS  CAS  Google Scholar 

  3. Clute, P. & Pines, J. Temporal and spatial control of cyclin B1 destruction in metaphase. Nature Cell Biol. 1, 82–87 (1999)

    Article  CAS  Google Scholar 

  4. Dick, A. E. & Gerlich, D. W. Kinetic framework of spindle assembly checkpoint signalling. Nature Cell Biol. 15, 1370–1377 (2013)

    Article  CAS  Google Scholar 

  5. Sudakin, V., Chan, G. K. & Yen, T. J. Checkpoint inhibition of the APC/C in HeLa cells is mediated by a complex of BUBR1, BUB3, CDC20, and MAD2. J. Cell Biol. 154, 925–936 (2001)

    Article  CAS  Google Scholar 

  6. Chao, W. C., Kulkarni, K., Zhang, Z., Kong, E. H. & Barford, D. Structure of the mitotic checkpoint complex. Nature 484, 208–213 (2012)

    Article  ADS  CAS  Google Scholar 

  7. Han, J. S. et al. Catalytic assembly of the mitotic checkpoint inhibitor BubR1-Cdc20 by a Mad2-induced functional switch in Cdc20. Mol. Cell 51, 92–104 (2013)

    Article  CAS  Google Scholar 

  8. Izawa, D. & Pines, J. Mad2 and the APC/C compete for the same site on Cdc20 to ensure proper chromosome segregation. J. Cell Biol. 199, 27–37 (2012)

    Article  CAS  Google Scholar 

  9. Primorac, I. & Musacchio, A. Panta rhei: the APC/C at steady state. J. Cell Biol. 201, 177–189 (2013)

    Article  CAS  Google Scholar 

  10. Izawa, D. & Pines, J. How APC/C-Cdc20 changes its substrate specificity in mitosis. Nature Cell Biol. 13, 223–233 (2011)

    Article  CAS  Google Scholar 

  11. Tang, Z., Bharadwaj, R., Li, B. & Yu, H. Mad2-independent inhibition of APCCdc20 by the mitotic checkpoint protein BubR1. Dev. Cell 1, 227–237 (2001)

    Article  CAS  Google Scholar 

  12. Burton, J. L. & Solomon, M. J. Mad3p, a pseudosubstrate inhibitor of APCCdc20 in the spindle assembly checkpoint. Genes Dev. 21, 655–667 (2007)

    Article  CAS  Google Scholar 

  13. King, E. M., van der Sar, S. J. & Hardwick, K. G. Mad3 KEN boxes mediate both Cdc20 and Mad3 turnover, and are critical for the spindle checkpoint. PLoS ONE 2, e342 (2007)

    Article  ADS  Google Scholar 

  14. Tian, W. et al. Structural analysis of human Cdc20 supports multisite degron recognition by APC/C. Proc. Natl Acad. Sci. USA 109, 18419–18424 (2012)

    Article  ADS  CAS  Google Scholar 

  15. Elowe, S. et al. Uncoupling of the spindle-checkpoint and chromosome-congression functions of BubR1. J. Cell Sci. 123, 84–94 (2010)

    Article  CAS  Google Scholar 

  16. Lara-Gonzalez, P., Scott, M. I., Diez, M., Sen, O. & Taylor, S. S. BubR1 blocks substrate recruitment to the APC/C in a KEN-box-dependent manner. J. Cell Sci. 124, 4332–4345 (2011)

    Article  CAS  Google Scholar 

  17. Collin, P., Nashchekina, O., Walker, R. & Pines, J. The spindle assembly checkpoint works like a rheostat rather than a toggle switch. Nature Cell Biol. 15, 1378–1385 (2013)

    Article  CAS  Google Scholar 

  18. Rothbauer, U. et al. A versatile nanotrap for biochemical and functional studies with fluorescent fusion proteins. Mol. Cell. Proteomics 7, 282–289 (2008)

    Article  CAS  Google Scholar 

  19. Lau, D. T. & Murray, A. W. Mad2 and Mad3 cooperate to arrest budding yeast in mitosis. Curr. Biol. 22, 180–190 (2012)

    Article  CAS  Google Scholar 

  20. Santaguida, S., Tighe, A., D'Alise, A. M., Taylor, S. S. & Musacchio, A. Dissecting the role of MPS1 in chromosome biorientation and the spindle checkpoint through the small molecule inhibitor reversine. J. Cell Biol. 190, 73–87 (2010)

    Article  CAS  Google Scholar 

  21. Kiyomitsu, T., Obuse, C. & Yanagida, M. Human blinkin/AF15q14 is required for chromosome alignment and the mitotic checkpoint through direct interaction with Bub1 and BubR1. Dev. Cell 13, 663–676 (2007)

    Article  CAS  Google Scholar 

  22. De Antoni, A. et al. The Mad1/Mad2 complex as a template for Mad2 activation in the spindle assembly checkpoint. Curr. Biol. 15, 214–225 (2005)

    Article  CAS  Google Scholar 

  23. Mariani, L. et al. Role of the Mad2 dimerization interface in the spindle assembly checkpoint independent of kinetochores. Curr. Biol. (2012)

  24. Tang, Z., Shu, H., Oncel, D., Chen, S. & Yu, H. Phosphorylation of Cdc20 by Bub1 provides a catalytic mechanism for APC/C inhibition by the spindle checkpoint. Mol. Cell 16, 387–397 (2004)

    Article  CAS  Google Scholar 

  25. Westhorpe, F. G., Tighe, A., Lara-Gonzalez, P. & Taylor, S. S. p31comet-mediated extraction of Mad2 from the MCC promotes efficient mitotic exit. J. Cell Sci. 124, 3905–3916 (2011)

    Article  CAS  Google Scholar 

  26. Varetti, G., Guida, C., Santaguida, S., Chiroli, E. & Musacchio, A. Homeostatic control of mitotic arrest. Mol. Cell 44, 710–720 (2011)

    Article  CAS  Google Scholar 

  27. Mansfeld, J., Collin, P., Collins, M. O., Choudhary, J. & Pines, J. APC15 drives the turnover of MCC-Cdc20 to make the spindle assembly checkpoint responsive to kinetochore attachment. Nature Cell Biol. 13, 1234–1243 (2011)

    Article  CAS  Google Scholar 

  28. Nilsson, J., Yekezare, M., Minshull, J. & Pines, J. The APC/C maintains the spindle assembly checkpoint by targeting Cdc20 for destruction. Nature Cell Biol. 10, 1411–1420 (2008)

    Article  CAS  Google Scholar 

  29. Matyskiela, M. E. & Morgan, D. O. Analysis of activator-binding sites on the APC/C supports a cooperative substrate-binding mechanism. Mol. Cell 34, 68–80 (2009)

    Article  CAS  Google Scholar 

Download references

Acknowledgements

We are grateful to T. Matsusaka for developing infrared-dye-conjugated ubiquitylation substrates, to A. Musacchio, W. Earnshaw, T. Kiyomitsu and M. Yanagida for reagents, and to A. Musacchio and members of our laboratory for critical discussions. This work was supported by a project and a programme grant from Cancer Research UK to J.P. J.P. acknowledges core funding to the Gurdon Institute from the Wellcome Trust and CR UK.

Author information

Authors and Affiliations

Authors

Contributions

Experiments were designed by D.I. and J.P., carried out by D.I., and analysed by D.I. and J.P.; D.I. and J.P. wrote the paper.

Corresponding author

Correspondence to Jonathon Pines.

Ethics declarations

Competing interests

The authors declare no competing financial interests.

Extended data figures and tables

Extended Data Figure 1 Recombinant human mitotic checkpoint complex binds to a second CDC20.

a, Schematic illustration of purification steps. Human wild-type CDC20 (untagged), SBPBUBR1 and 6HisMAD2 were expressed in baculovirus-infected Sf9 cells. The recombinant core mitotic checkpoint complex (rMCC) was purified by nitrilotriacetic acid (Ni-NTA) and streptavidin beads. Purified core rMCC bound to streptavidin beads was used to assay binding to purified recombinant Cdc20. b, Core rMCC consisting of CDC20, SBPBUBR1 and 6HisMAD2 was analysed by SDS–PAGE and Coomassie blue R250 staining, followed by quantification at 680 nm on a LiCOR Odyssey scanner. Equal molar amounts of purified SBPBUBR1, SBPCDC20 and SBPMAD2 proteins were used to calibrate the Coomassie blue staining. The stoichiometry of core rMCC (mean ± s.d. is shown below the panel with SBPBUBR1 set to 1.0) was estimated from three independently purified core rMCC preparations. Molecular mass markers are on the left. c, d, Both the MAD2 binding motif of CDC20 and the first KEN box of BUBR1 are required to assemble rMCC. Core rMCC was pulled down with streptavidin beads from Sf9 cells expressing SBPBUBR1, 6HisMAD2 and either wild-type (WT) CDC20 or the K129ILR/AAAA mutant (ΔKILR) (c), or 6HisMAD2, wild-type CDC20 plus wild-type SBPBUBR1, or alanine substitution mutants of either KEN box 1 (ΔKEN1) or KEN box 2 (ΔKEN2). The proteins retained on streptavidin beads were analysed by immunoblotting with the indicated antibodies. e, Relative expression levels of core rMCC components. Sf9 cells extracts expressing the core rMCC, and the purified rMCC complex, were analysed by quantitative immunoblotting. The ratio of the proteins in the extracts is given, with that of SBPBUBR1 set to 1.0. f, Schematic illustration of the second CDC20 binding assay in Fig. 1a. In lanes 1 and 2, the streptavidin beads were incubated with either 6HisCDC20 wild-type or the ΔKILR (K129ILR/AAAA) mutant. In lanes 3 and 4, the streptavidin beads bound to core rMCC were incubated with the 6HisCDC20 proteins. In lanes 6 and 7, the streptavidin beads bound to 6His-SBPCDC20 were incubated with the 6HisCDC20 proteins. g, Sf9 cell extracts expressing core rMCC or 3Flag-tagged CDC20 were mixed and the core rMCC purified as in a. The core rMCC was analysed by quantitative immunoblotting. 51% of the core rMCC was purified bound to a second 3FlagCDC20. h, A functional CDC20 promotes the binding of core rMCC to the APC/C. The APC/C was immunoprecipitated from CDC20-depleted mitotic extracts supplemented with a constant amount of core rMCC and tenfold excess of recombinant wild-type SBPCDC20, or the ΔKILR or ΔIR mutants. The co-immunoprecipitates were analysed as in Fig. 1c. i, Schematic of the APC/C–MCC–CDC20 ternary complex. Both core rMCC and CDC20 bind to the APC/C and form a ternary complex (left). The CDC20ΔKILR mutant cannot bind the APC/C directly, nor stimulate core rMCC binding to the APC/C, but CDC20ΔKILR still binds to rMCC (right). All results are representative of two or more independent biological replicates.

Extended Data Figure 2 Comparison of rMCC with and without BUB3.

a, b, Preparation of recombinant core MCC with or without BUB3. Insect cells were infected with viruses expressing core MCC components with and without BUB3, and the rMCC was purified by Ni-NTA and streptavidin beads. The complexes were analysed by Coomassie blue (CB) staining (a) and immunoblotting (b). c, Binding to a second 6HisCDC20 of recombinant core MCC with or without BUB3 was performed and analysed as in Fig. 1a. All results are representative of two independent biological replicates.

Extended Data Figure 3 Molar ratios of rMCC, CDC20 and the APC/C in the in vitro ubiquitylation assays.

a, b, Core rMCC and CDC20 from Fig. 1d were analysed by quantitative immunoblotting. CDC20, MAD2 and BUBR1 were analysed by quantitative immunoblotting in the input (a) and in the reaction (b). The black filled circles are unconjugated SBPCDC20; red filled circles are ubiquitylated SBPCDC20. c, Core rMCC, rCDC20, and the APC/C immunoprecipitates used in Fig. 1e, plus a purified SBPAPC3 subunit, were analysed by quantitative immunoblotting with the indicated antibodies. The calculated molar ratios of rMCC, rCDC20 and the APC/C are shown below the panels.

Extended Data Figure 4 Cells expressing the D-box and KEN box receptor mutants of CDC20 can degrade cyclin B1 in nocodazole.

Cyclin B1–Venus degradation was analysed in siRNA CDC20-treated cells rescued with siRNA-resistant versions of 3×Flag–CDC20, wild-type, or ΔDR, or ΔKR mutants, in the presence of nocodazole (0.33 µM). The fluorescence of individual cells was measured, the value at NEBD set to 1 and the mean ± s.e.m. for all cells plotted. n, number of cells analysed from at least two independent experiments.

Extended Data Figure 5 Characterization of the MCC containing D-box or KEN box 2 mutants of BUBR1.

a, Core rMCC assembled with SBPBUBR1 wild type, or ΔD-box, or ΔKEN2 mutants, was purified as in Extended Data Fig. 1a, b and analysed on a LiCOR Odyssey scanner at 680 nm after SDS–PAGE and Coomassie blue R250 staining. b, The core rMCC mutants prepared in a were assayed as APC/C inhibitors in an in vitro ubiquitylation assay as in Fig. 1d. c, Insect cell extracts expressing CDC20 with SBPBUBR1, either wild type, or ΔD-box or ΔKEN2 mutants, were incubated with streptavidin beads. The proteins retained on the streptavidin beads were analysed by quantitative immunoblotting. Results in panels ac are representative of two independent biological replicates. d, HeLa cells were treated with siRNA against BUBR1 and rescued with 3×Flag–Cerulean–BUBR1, either wild-type or the ΔD-box mutant, and mitosis analysed in 0.116 µM Taxol as in Fig. 3d. The time from NEBD to anaphase (or mitotic exit) was measured and plotted as a box and whisker chart. n, number of analysed cells from two independent biological replicates. e, HeLa cells were treated with siRNA against BUBR1 and rescued with siRNA resistant 3×Flag–Cerulean–BUBR1, either wild type or the ΔD-box mutant, then analysed by immunostaining. Cells were stained with anti-Flag M2 and anti-ACA antibodies, and Hoechst 33342, and representative images of prometaphase cells from two independent biological replicates are shown. Scale bar, 10 µm.

Extended Data Figure 6 Stabilizing the interaction between MAD2 and CDC20.

a, Schematic of how a stabilized MCC might block cells in metaphase. At prometaphase, when the SAC is ‘ON’, CDC20 is inhibited both by incorporation into the MCC and through binding to the MCC. At metaphase when the SAC is ‘OFF’, CDC20 is released from the MCC and activates the APC/C. We postulate that stabilizing an exogenous MCC to prevent its disassembly should also prevent endogenous CDC20 from activating the APC/C, which results in an anaphase delay. b, Schematic of a stabilized MCC. To stabilize the MCC we took advantage of the binding between yellow fluorescent protein (Venus) and GFP-binding domain (GBP), which is a 13kDa domain from a camelid antibody that binds strongly and specifically to GFP and YFP18. MAD2 and BUBR1 were tagged with Venus and the GBP domain was tagged to CDC20. We refer to the MCC containing a stabilized MAD2–CDC20 interaction as MCCM2, and that with stabilized BUBR1–CDC20 as MCCR1. c, GBP- and Venus-fusion proteins bind stably to each other in vivo. HeLa cell lines expressing siRNA-resistant myc–CDC20 or myc–GBP–CDC20 were transfected with plasmids encoding either Venus alone or Venus–MAD2, followed by siRNA treatment against CDC20. After a single thymidine block and release, the cells were arrested at prometaphase by treating with nocodazole, and harvested by mitotic shake-off 48 h after the siRNA treatment. Proteins were immunoprecipitated with anti-myc epitope antibodies before analysis by quantitative immunoblotting with the indicated antibodies. WT, myc–CDC20; GBP, myc–GBP–CDC20. Results in panel c are representative of three independent biological replicates.

Extended Data Figure 7 Stabilizing the interaction between MAD2 and CDC20 prevents disassembly of the MCC in vivo.

a, b, Tethering CDC20 to MAD2 prevents MCC disassembly and release from the APC/C. a, Empty plasmids or plasmids encoding Venus–MAD2 were transfected into HeLa cell lines expressing 3×Flag–GBP–CDC20 and the cells synchronized at prometaphase by thymidine release followed by a nocodazole block. Cells were harvested by mitotic shake off and separated into two cultures after washing once in medium. One culture was harvested immediately (−reversine) and the other resuspended in medium containing 1 µM reversine and 10 µM MG132 (+reversine) for 1 h before harvesting. The APC/C was immunoprecipitated with an anti-APC4 antibody and the immunoprecipitates analysed by quantitative immunoblotting. We note that the APC/C preferred to bind endogenous CDC20 over GBP–CDC20 as the co-activator in vivo (see +reversine lane in control cells) but the MCCM2 did not sequester endogenous CDC20 from the APC/C (see +reversine lane in GBP–CDC20 + Venus–MAD2 cells). b, Mean ± s.e.m. of the relative amounts of the indicated proteins in the APC4 immunoprecipitates calculated from four independent biological experiments. The amount of protein bound to the APC/C in the absence of reversine was set to 1 (red line). ce, Tethering CDC20 to MAD2 prevents MCC disassembly and release from the APC/C in the absence of endogenous CDC20. c, Plasmids encoding Venus–MAD2 were transfected into HeLa cell lines expressing the indicated CDC20 fusion proteins following siRNA treatment against CDC20 for 48 h. Cells were synchronized at prometaphase then treated with reversine, and anti APC4 and anti-GFP immunoprecipitates were analysed as in a. WT, myc–CDC20; GBP, myc–GBP–CDC20. Note that endogenous CDC20 could not be inhibited through exchange into MCCM2 because a core MCC composed of Venus–MAD2 and untagged CDC20 disassembled. d, HeLa cell lines expressing myc–CDC20 (upper blots) or myc–GBP–CDC20 (lower blots) were transfected with a plasmid encoding Venus–MAD2 followed by siRNA treatment against CDC20 for 48 h. Cells were synchronized at prometaphase and treated with reversine as indicated in a. Total cell extracts were analysed by size exclusion chromatography on a Sepharose 6 column and fractions were analysed by quantitative immunoblotting against the indicated proteins and the relative amounts of Venus–MAD2 plotted in panel e with the sum of Venus–MAD2 intensities set to 1. The migration of APC/C or APC/C-MCC is annotated below panel d. All results are representative of three independent biological replicates.

Extended Data Figure 8 KNL1 (also known as CASC5) is not required for a stabilized MCC to inhibit anaphase.

a, HeLa cells expressing MCCM2 in Fig. 4a were analysed by immunostaining. The cells were stained with anti-GFP, anti-MAD2, anti-ACA and Hoechst 33342, and representative images of prometaphase and metaphase cells from two independent biological replicates are shown. Scale bar, 5 µm. b, The time from NEBD to anaphase in Fig. 4a was plotted against the intensity of mCherry–GBP–CDC20 (left) or the ratio of Venus–MAD2 to mCherry–GBP–CDC20 (right). The ratio of Venus–MAD2 to mCherry–GBP–CDC20 was calibrated by measuring fluorescence intensity of a mCherry–GBP–Venus fusion protein in HeLa cells. ce, MCCM2 delays anaphase when KNL1 is depleted. c, HeLa cells were treated with siRNA against KNL1 for 72 h and total cell extracts were analysed by quantitative immunoblotting with the indicated antibodies. d, HeLa cells treated as in c were analysed by immunostaining. The cells were stained with anti-CDC20, anti-BUBR1, anti-ACA and Hoechst 33342, and representative images of early prometaphase from two independent biological replicates are shown. Scale bar, 10 µm. e, HeLa cell lines stably expressing mCherry–GBP–CDC20 and an inducible 3×Flag–Venus–MAD2 (expressed from a tetracyclin-inducible promoter) were treated with siRNA against KNL1 as in c. Progression through mitosis was analysed in the presence (+Tet) or absence (−Tet) of tetracyclin, and analysed as in Fig. 4b. n, number of cells from two independent biological replicates.

Extended Data Figure 9 Functional MCCM2 is required to delay anaphase.

a, HeLa cells were transfected with plasmids encoding Venus–MAD2 and either wild-type or a MAD2-binding defective (ΔKILR) mutant of CDC20 tagged with mCherry–GBP, and mitotic progression was analysed as in Fig. 4a. n, number of cells from three independent biological replicates. b, The core rMCC mutants used in Extended Data Fig. 5a were incubated with preformed APC/CCDC20 and assayed as APC/C inhibitors in an in vitro ubiquitylation assay as in Fig. 1e. The extent of APC/C inhibition (incubation of MCCWT set to 1.0) is shown below the securin panel. This result is representative of two independent experiments. c, Schematic of the inhibitory activities of the stabilized MCCs in BUBR1-depleted cells used in Fig. 4c. When BUBR1 is depleted, MAD2 and CDC20 cannot form the MCC to inhibit endogenous CDC20 (left). When rescued with wild-type BUBR1, MCCM2 can form and inhibit endogenous CDC20 to delay anaphase. By contrast, when rescued by the BUBR1 ΔD-box mutant, MCCM2 can only weakly inhibit endogenous CDC20 and cells can proceed into anaphase.

Extended Data Figure 10 Model for how the MCC could disseminate the ‘wait anaphase’ signal.

Unattached kinetochores catalyse MCC formation and the MCC disseminates the ‘wait anaphase’ signal through the cytoplasm (black arrows). When the MCC disassembles (blue arrows), this releases CDC20, which along with newly synthesized CDC20, can have two fates: to be recruited to unattached kinetochores and incorporated into the MCC, or to bind the APC/C to form APC/CCDC20. The MCC is able to inhibit both unbound CDC20 and CDC20 bound to the APC/C (red bars).

PowerPoint slides

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Izawa, D., Pines, J. The mitotic checkpoint complex binds a second CDC20 to inhibit active APC/C. Nature 517, 631–634 (2015). https://doi.org/10.1038/nature13911

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/nature13911

This article is cited by

Comments

By submitting a comment you agree to abide by our Terms and Community Guidelines. If you find something abusive or that does not comply with our terms or guidelines please flag it as inappropriate.

Search

Quick links

Nature Briefing

Sign up for the Nature Briefing newsletter — what matters in science, free to your inbox daily.

Get the most important science stories of the day, free in your inbox. Sign up for Nature Briefing