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.

  • Article
  • Published:

Condensin confers the longitudinal rigidity of chromosomes

Nature Cell Biology volume 17, pages 771–781 (2015)Cite this article

Subjects

Abstract

In addition to inter-chromatid cohesion, mitotic and meiotic chromatids must have three physical properties: compaction into ‘threads’ roughly co-linear with their DNA sequence, intra-chromatid cohesion determining their rigidity, and a mechanism to promote sister chromatid disentanglement. A fundamental issue in chromosome biology is whether a single molecular process accounts for all three features. There is universal agreement that a pair of Smc–kleisin complexes called condensin I and II facilitate sister chromatid disentanglement, but whether they also confer thread formation or longitudinal rigidity is either controversial or has never been directly addressed respectively. We show here that condensin II (beta-kleisin) has an essential role in all three processes during meiosis I in mouse oocytes and that its function overlaps with that of condensin I (gamma-kleisin), which is otherwise redundant. Pre-assembled meiotic bivalents unravel when condensin is inactivated by TEV cleavage, proving that it actually holds chromatin fibres together.

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

Access options

Buy this article

  • Purchase on Springer Link
  • Instant access to full article PDF
Buy now

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

Figure 1: Condensin II but not condensin I is essential for meiosis.
Figure 2: Condensin II determines the morphology and rigidity of meiotic bivalents.
Figure 3: Deletion of Ncaph1 or Ncaph2 induces different condensation defects.
Figure 4: Condensin is required for sister chromatid disentanglement even in the absence of cohesin.
Figure 5: Condensins I and II have overlapping functions.
Figure 6: Creation of a functional TEV-cleavable Ncaph2.
Figure 7: Condensin maintains the morphology and rigidity of chromosomes.
Figure 8: The stable association of condensin II with chromosomes maintains their structure.

Similar content being viewed by others

References

  1. Cremer, T., Tesin, D., Hopman, A. H. & Manuelidis, L. Rapid interphase and metaphase assessment of specific chromosomal changes in neuroectodermal tumor cells by in situ hybridization with chemically modified DNA probes. Exp. Cell Res. 176, 199–220 (1988).

    Article  CAS  Google Scholar 

  2. Flemming, W. Beitrage zur Kenntnisseder Zelle undIhreer Lebenserscheinungen. Archiv f. mikrosk. Anatomie. 18, 151–259 (1880).

    Article  Google Scholar 

  3. Nasmyth, K. Segregating sister genomes: the molecular biology of chromosome separation. Science 297, 559–565 (2002).

    Article  CAS  Google Scholar 

  4. Nasmyth, K. Disseminating the genome: joining, resolving, and separating sister chromatids during mitosis and meiosis. Annu. Rev. Genet. 35, 673–745 (2001).

    Article  CAS  Google Scholar 

  5. Schleiffer, A. et al. Kleisins: a superfamily of bacterial and eukaryotic SMC protein partners. Mol. Cell 11, 571–575 (2003).

    Article  CAS  Google Scholar 

  6. Hirano, T. & Mitchison, T. J. A heterodimeric coiled-coil protein required for mitotic chromosome condensation in vitro. Cell 79, 449–458 (1994).

    Article  CAS  Google Scholar 

  7. Hirano, T., Kobayashi, R. & Hirano, M. Condensins, chromosome condensation protein complexes containing XCAP-C, XCAP-E and a Xenopus homolog of the Drosophila Barren protein. Cell 89, 511–521 (1997).

    Article  CAS  Google Scholar 

  8. Ono, T. et al. Differential contributions of condensin I and condensin II to mitotic chromosome architecture in vertebrate cells. Cell 115, 109–121 (2003).

    Article  CAS  Google Scholar 

  9. Saka, Y. et al. Fission yeast cut3 and cut14, members of a ubiquitous protein family, are required for chromosome condensation and segregation in mitosis. EMBO J. 13, 4938–4952 (1994).

    Article  CAS  Google Scholar 

  10. Sutani, T. et al. Fission yeast condensin complex: essential roles of non-SMC subunits for condensation and Cdc2 phosphorylation of Cut3/SMC4. Genes Dev. 13, 2271–2283 (1999).

    Article  CAS  Google Scholar 

  11. Strunnikov, A. V., Hogan, E. & Koshland, D. SMC2, a Saccharomyces cerevisiae gene essential for chromosome segregation and condensation, defines a subgroup within the SMC family. Genes Dev. 9, 587–599 (1995).

    Article  CAS  Google Scholar 

  12. Lavoie, B. D., Tuffo, K. M., Oh, S., Koshland, D. & Holm, C. Mitotic chromosome condensation requires Brn1p, the yeast homologue of Barren. Mol. Biol. Cell 11, 1293–1304 (2000).

    Article  CAS  Google Scholar 

  13. Bhalla, N., Biggins, S. & Murray, A. W. Mutation of YCS4, a budding yeast condensin subunit, affects mitotic and nonmitotic chromosome behavior. Mol. Biol. Cell 13, 632–645 (2002).

    Article  CAS  Google Scholar 

  14. Lieb, J. D., Albrecht, M. R., Chuang, P. T. & Meyer, B. J. MIX-1: an essential component of the C. elegans mitotic machinery executes X chromosome dosage compensation. Cell 92, 265–277 (1998).

    Article  CAS  Google Scholar 

  15. Hagstrom, K. A., Holmes, V. F., Cozzarelli, N. R. & Meyer, B. J. C. elegans condensin promotes mitotic chromosome architecture, centromere organization, and sister chromatid segregation during mitosis and meiosis. Genes Dev. 16, 729–742 (2002).

    Article  CAS  Google Scholar 

  16. Bhat, M. A., Philp, A. V., Glover, D. M. & Bellen, H. J. Chromatid segregation at anaphase requires the barren product, a novel chromosome-associated protein that interacts with Topoisomerase II. Cell 87, 1103–1114 (1996).

    Article  Google Scholar 

  17. Steffensen, S. et al. A role for Drosophila SMC4 in the resolution of sister chromatids in mitosis. Curr. Biol. 11, 295–307 (2001).

    Article  CAS  Google Scholar 

  18. Hudson, D. F., Vagnarelli, P., Gassmann, R. & Earnshaw, W. C. Condensin is required for nonhistone protein assembly and structural integrity of vertebrate mitotic chromosomes. Dev. Cell 5, 323–336 (2003).

    Article  CAS  Google Scholar 

  19. Vagnarelli, P. et al. Condensin and Repo-Man-PP1 co-operate in the regulation of chromosome architecture during mitosis. Nat. Cell Biol. 8, 1133–1142 (2006).

    Article  CAS  Google Scholar 

  20. Gerlich, D., Hirota, T., Koch, B., Peters, J. M. & Ellenberg, J. Condensin I stabilizes chromosomes mechanically through a dynamic interaction in live cells. Curr. Biol. 16, 333–344 (2006).

    Article  CAS  Google Scholar 

  21. Ribeiro, S. A. et al. Condensin regulates the stiffness of vertebrate centromeres. Mol. Biol. Cell 20, 2371–2380 (2009).

    Article  CAS  Google Scholar 

  22. Oliveira, R. A., Coelho, P. A. & Sunkel, C. E. The condensin I subunit Barren/CAP-H is essential for the structural integrity of centromeric heterochromatin during mitosis. Mol. Cell Biol. 25, 8971–8984 (2005).

    Article  CAS  Google Scholar 

  23. Poirier, M. G. & Marko, J. F. Micromechanical studies of mitotic chromosomes. Curr. Top Dev. Biol. 55, 75–141 (2003).

    Article  CAS  Google Scholar 

  24. Almagro, S., Riveline, D., Hirano, T., Houchmandzadeh, B. & Dimitrov, S. The mitotic chromosome is an assembly of rigid elastic axes organized by structural maintenance of chromosomes (SMC) proteins and surrounded by a soft chromatin envelope. J. Biol. Chem. 279, 5118–5126 (2004).

    Article  CAS  Google Scholar 

  25. Nicklas, R. B. How cells get the right chromosomes. Science 275, 632–637 (1997).

    Article  CAS  Google Scholar 

  26. Lan, Z. J., Xu, X. & Cooney, A. J. Differential oocyte-specific expression of Cre recombinase activity in GDF-9-iCre, Zp3cre, and Msx2Cre transgenic mice. Biol. Reprod. 71, 1469–1474 (2004).

    Article  CAS  Google Scholar 

  27. Sironi, L. et al. Mad2 binding to Mad1 and Cdc20, rather than oligomerization, is required for the spindle checkpoint. EMBO J. 20, 6371–6382 (2001).

    Article  CAS  Google Scholar 

  28. Miyanari, Y., Ziegler-Birling, C. & Torres-Padilla, M. E. Live visualization of chromatin dynamics with fluorescent TALEs. Nat. Struct. Mol. Biol. 20, 1321–1324 (2013).

    Article  CAS  Google Scholar 

  29. Lee, J., Ogushi, S., Saitou, M. & Hirano, T. Condensins I and II are essential for construction of bivalent chromosomes in mouse oocytes. Mol. Biol. Cell 22, 3465–3477 (2011).

    Article  CAS  Google Scholar 

  30. Tachibana-Konwalski, K. et al. Rec8-containing cohesin maintains bivalents without turnover during the growing phase of mouse oocytes. Genes Dev. 24, 2505–2516 (2010).

    Article  CAS  Google Scholar 

  31. Wassmann, K., Niault, T. & Maro, B. Metaphase I arrest upon activation of the Mad2-dependent spindle checkpoint in mouse oocytes. Curr. Biol. 13, 1596–1608 (2003).

    Article  CAS  Google Scholar 

  32. Hirota, T., Gerlich, D., Koch, B., Ellenberg, J. & Peters, J. M. Distinct functions of condensin I and II in mitotic chromosome assembly. J. Cell Sci. 117, 6435–6445 (2004).

    Article  CAS  Google Scholar 

  33. Renshaw, M. J. et al. Condensins promote chromosome recoiling during early anaphase to complete sister chromatid separation. Dev. Cell 19, 232–244 (2010).

    Article  CAS  Google Scholar 

  34. Charbin, A., Bouchoux, C. & Uhlmann, F. Condensin aids sister chromatid decatenation by topoisomerase II. Nucleic Acids Res. 42, 340–348 (2014).

    Article  CAS  Google Scholar 

  35. Poirier, M., Eroglu, S., Chatenay, D. & Marko, J. F. Reversible and irreversible unfolding of mitotic newt chromosomes by applied force. Mol. Biol. Cell 11, 269–276 (2000).

    Article  CAS  Google Scholar 

  36. Cuylen, S., Metz, J. & Haering, C. H. Condensin structures chromosomal DNA through topological links. Nat. Struct. Mol. Biol. 18, 894–901 (2011).

    Article  CAS  Google Scholar 

  37. Shintomi, K. & Hirano, T. The relative ratio of condensin I to II determines chromosome shapes. Genes Dev. 25, 1464–1469 (2011).

    Article  CAS  Google Scholar 

  38. Fujiwara, T., Tanaka, K., Kuroiwa, T. & Hirano, T. Spatiotemporal dynamics of condensins I and II: evolutionary insights from the primitive red alga Cyanidioschyzon merolae. Mol. Biol. Cell 24, 2515–2527 (2013).

    Article  CAS  Google Scholar 

  39. Burmann, F. et al. An asymmetric SMC-kleisin bridge in prokaryotic condensin. Nat. Struct. Mol. Biol. 20, 371–379 (2013).

    Article  Google Scholar 

  40. Cuylen, S., Metz, J., Hruby, A. & Haering, C. H. Entrapment of chromosomes by condensin rings prevents their breakage during cytokinesis. Dev. Cell 27, 469–478 (2013).

    Article  CAS  Google Scholar 

  41. Oliveira, R. A., Hamilton, R. S., Pauli, A., Davis, I. & Nasmyth, K. Cohesin cleavage and Cdk inhibition trigger formation of daughter nuclei. Nat. Cell Biol. 12, 185–192 (2010).

    Article  CAS  Google Scholar 

  42. Nagy, A., Gertsenstein, M., Vintersten, K. & Behringer, R. Manipulating the Mouse Embryo A Laboratory Manual 3rd edn (Cold Spring Harbor Laboratory Press, 2003).

    Google Scholar 

  43. Kitajima, T. S., Ohsugi, M. & Ellenberg, J. Complete kinetochore tracking reveals error-prone homologous chromosome biorientation in mammalian oocytes. Cell 146, 568–581 (2011).

    Article  CAS  Google Scholar 

  44. Rattani, A. et al. Sgol2 provides a regulatory platform that coordinates essential cell cycle processes during meiosis I in oocytes. eLife 2, e01133 (2013).

    Article  Google Scholar 

Download references

Acknowledgements

We thank Micron microscopy facility and especially I. Dobbie for his technical support; all the staff of the BSB facility; B. Novák and all the members of the K.N. laboratory for discussions and comments on the manuscript. This work was supported by the Wellcome Trust (Grant Ref 091859/Z/10/Z), ERC grant (Proposal No 294401) and MRC (Grant ID 84673).

Author information

Authors and Affiliations

  1. Department of Biochemistry, University of Oxford, South Parks Road, Oxford OX1 3QU, UK

    Martin Houlard, Jonathan Godwin, Jean Metson & Kim Nasmyth

  2. Laboratory of Developmental Biotechnology, Graduate School of Agricultural Science, Kobe University, 1-1 Rokkodai-cho, Nada-ku, Kobe 657-8501, Japan

    Jibak Lee

  3. Chromosome Dynamics Laboratory, RIKEN, 2-1 Hirosawa, Wako, Saitama 351-0198, Japan

    Tatsuya Hirano

Authors
  1. Martin Houlard
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Jonathan Godwin
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Jean Metson
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Jibak Lee
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Tatsuya Hirano
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Kim Nasmyth
    View author publications

    You can also search for this author in PubMed Google Scholar

Contributions

M.H. designed, performed and analysed the experiments. J.G. provided technical advice for oocytes manipulation. J.M. realized all the genotyping. J.L. provided reagents. T.H. provided reagents. K.N. designed and analysed the experiments and coordinated the work.

Corresponding author

Correspondence to Kim Nasmyth.

Ethics declarations

Competing interests

The authors declare no competing financial interests.

Integrated supplementary information

Supplementary Figure 1

(A) Ncaph1 is not essential during meiosis I. Oocytes from Ncaph1f/f females (Wild type Ncaph1) or Ncaph1f/f Tg(ZP3Cre) females (ΔNcaph1) were injected at GV stage with mRNA coding for H2B-mCherry to mark the whole chromosomes in red and TALE-mClover_MajSat to mark the pericentric repeats in green. Meiosis I progression was followed by live cell confocal imaging. Maximum intensity z projection images of the major time points are shown. The phenotype was observed in all oocytes analysed (24 oocytes from 3 females in 3 experiments). Scale bar: 10 μm. (B) Segregation defects inΔNcaph2 oocytes Snapshot 16 h post GVBD of one of the 18% of ΔNcaph2 oocytes that go through polar body extrusion. Phenotype observed in all the 18% of ΔNcaph2 oocytes that extrude a polar body. Scale bar, 10 μm.

Supplementary Figure 2 Deletion of Ncaph1 doesn’t impede on Ncaph2 loading.

Chromosomes from oocytes isolated from Ncaph1f/fTg(ZP3Cre) or Ncaph2f/f Tg(ZP3Cre) females were analysed by chromosome spreading 7 h (meiosis I) or 16 h (meiosis II) after GVBD. Immunofluorescence was performed using antibodies directed against NCAPH2 and NCAPH1 on ΔNcaph1 (A) and ΔNcaph2 oocytes (B) respectively. The DNA was stained with DAPI. Oocytes from females Ncaph1f/f or Ncaph2f/f were used as control (Wild type). The phenotypes were observed in all oocytes analysed analysed (More than 10 oocytes from 2 females in 2 experiments). Scale bar, 5 μm.

Supplementary Figure 3 Topoisomerase 2 alpha localization on ΔNcaph1 and ΔNcaph2 chromosome spreads.

(A) Chromosomes from oocytes isolated from Ncaph1f/f Tg(ZP3Cre) females were analysed by chromosome spreading 17 h (meiosis II) after GVBD. Immunofluorescence was performed using antibodies directed against Topoisomerase 2 Alpha. The DNA was stained with DAPI. Oocytes from Ncaph1f/f females were used as control (Wild type). The phenotype was observed in all oocytes analysed (14 oocytes from 2 females in 2 experiments). Scale bar: 5 μm. (B) Chromosomes from oocytes isolated from Ncaph2f/f Tg(ZP3Cre) females were analysed by chromosome spreading 7 h (meiosis I) or 17 h (meiosis II) after GVBD. Immunofluorescence was performed using antibodies directed against Topoisomerase 2 Alpha. The DNA was stained with DAPI. Oocytes from Ncaph2f/f females were used as control (Wild type). The phenotype was observed in all oocytes analysed (More than 5 oocytes from 2 females in 2 experimetns) Scale bar: 5 μm.

Supplementary Figure 4 GVBD kinetics of ΔNcaph1ΔNcaph2 oocytes.

Oocytes from Ncaph1f/f Ncaph2f/f and Ncaph1f/fNcaph2f/f Tg(ZP3Cre) females were isolated in M16 supplemented with IBMX. The kinetics starts as soon as the oocytes are washed in M16 to resume meiosis. The two groups of oocytes were followed by Live cell confocal microscopy to evaluate the time of GVBD. Results from two independent experiments performed using oocytes from one female of each genotype per experiment. The total number of oocytes are Ncaph1f/f Ncaph2f/f: n = 11 and Ncaph1f/f Ncaph2f/f Tg(ZP3Cre): n = 15. Error bars represent mean ± s.d.

Supplementary Figure 5 Chromosome spread analysis of NCAPH2-eGFP rescued Ncaph2f/f Tg(ZP3Cre) oocytes.

Oocytes from Ncaph2f/f Tg(ZP3Cre) females were injected at GV stage with Ncaph2-eGFP encoding RNA and analysed by chromosome spreading at 7 h (Meiosis I) and 16 h (Meiosis II) post GVBD using an antibody directed against eGFP. The phenotype was observed in all oocytes analysed (9 oocytes from 2 females in 2 experiments). Scale bar: 5 μm.

Supplementary information

Supplementary Information

Supplementary Information (PDF 1327 kb)

Meiosis I in Ncaph1 wild type oocytes.

Oocytes from Ncaph1f/f females were isolated in M16 supplemented with IBMX and injected at GV stage by mRNA encoding mCherry tagged H2B histone and TALE-mClover_MajSat. After two hours of incubation to allow the expression of the injected mRNA, IBXM was washed out and oocytes followed by live cell confocal microscopy. The time (h:min:s:ms) is indicated (top right). (MOV 25068 kb)

Meiosis I in oocytes deleted for Ncaph1.

Oocytes from Ncaph1f/f Tg(ZP3Cre) females were isolated in M16 supplemented with IBMX and injected at GV stage by mRNA encoding mCherry tagged H2B histone and TALE-mClover_MajSat. After two hours of incubation to allow the expression of the injected mRNA, IBXM was washed out and oocytes followed by live cell confocal microscopy. The time (h:min:s:ms) is indicated (top right). (MOV 26065 kb)

Meiosis I in Ncaph2 wild type oocytes.

Oocytes from Ncaph2f/f females were isolated in M16 supplemented with IBMX and injected at GV stage by mRNA encoding mCherry tagged H2B histone and TALE-mClover_MajSat. After two hours of incubation to allow the expression of the injected mRNA, IBXM was washed out and oocytes followed by live cell confocal microscopy. The time (h:min:s:ms) is indicated (top right). (MOV 15781 kb)

Meiosis I in oocytes deleted for Ncaph2.

Oocytes from Ncaph2f/f Tg(ZP3Cre) females were isolated in M16 supplemented with IBMX and injected at GV stage by mRNA encoding mCherry tagged H2B histone and TALE-mClover_MajSat. After two hours of incubation to allow the expression of the injected mRNA, IBXM was washed out and oocytes followed by live cell confocal microscopy. The time (h:min:s:ms) is indicated (top right). (MOV 34800 kb)

Meiosis I in oocytes Rec8Tev/Tev wild type Ncaph2.

Oocytes from Ncaph2f/f Rec8Tev/Tev females were isolated in M16 supplemented with IBMX and injected at GV stage by mRNA encoding mCherry tagged H2B histone, eGFP-Tubulin and TEV protease. After two hours of incubation to allow the expression of the injected mRNA, IBXM was washed out and oocytes followed by live cell confocal microscopy. The time (h:min:s:ms) is indicated (top right). (MOV 11414 kb)

Meiosis I in oocytes Rec8Tev/TevΔit Ncaph2.

Oocytes from Ncaph2f/f Tg(ZP3Cre) Rec8Tev/Tev females were isolated in M16 supplemented with IBMX and injected at GV stage by mRNA encoding mCherry tagged H2B histone, eGFP-Tubulin and TEV protease. After two hours of incubation to allow the expression of the injected mRNA, IBXM was washed out and oocytes followed by live cell confocal microscopy. The time (h:min:s:ms) is indicated (top right). (MOV 8065 kb)

Meiosis I in wild type Ncaph1 and 2 oocytes.

Oocytes from Ncaph1f/f Ncaph2f/f females were isolated in M16 supplemented with IBMX and injected at GV stage by mRNA encoding mCherry tagged H2B histone and TALE-mClover_MajSat. After two hours of incubation to allow the expression of the injected mRNA, IBXM was washed out and oocytes followed by live cell confocal microscopy. The time (h:min:s:ms) is indicated (top right). (MOV 16524 kb)

Meiosis I in oocytes deleted for Ncaph1 and Ncaph2.

Oocytes from Ncaph1f/f Ncaph2f/f Tg(ZP3Cre) females were isolated in M16 supplemented with IBMX and injected at GV stage by mRNA encoding mCherry tagged H2B histone and TALE-mClover_MajSat. After two hours of incubation to allow the expression of the injected mRNA, IBXM was washed out and oocytes followed by live cell confocal microscopy. The time (h:min:s:ms) is indicated (top right). (MOV 9072 kb)

In metaphase I, the chromosome shape is not altered by TEV protease in Δ Ncaph2 oocytes rescued by wild type Ncaph2.

Oocytes from Ncaph2f/f Tg(ZP3Cre) females were isolated in M16 supplemented with IBMX and injected at GV stage by mRNA encoding mCherry tagged H2B histone, TALE-mClover_MajSat, WT-NCAPH2 and MAD2. After two hours of incubation to allow the expression of the injected mRNA, IBXM was washed out and oocytes incubated overnight. Metaphase I arrested oocytes were then injected for a second time, with mRNA encoding TEV protease and followed by live cell confocal microscopy. The time (h:min:s:ms) is indicated (top right). (MOV 3023 kb)

In metaphase I, TEV protease induces the rapid stretching of chromosomes in Δ Ncaph2 oocytes rescued by TEV-Ncaph2.

Oocytes from Ncaph2f/f Tg(ZP3Cre) females were isolated in M16 supplemented with IBMX and injected at GV stage by mRNA encoding mCherry tagged H2B histone, TALE-mClover_MajSat, TEV-NCAPH2 and MAD2. After two hours of incubation to allow the expression of the injected mRNA, IBXM was washed out and oocytes incubated overnight. Metaphase I arrested oocytes were then injected for a second time, with mRNA encoding TEV protease and followed by live cell confocal microscopy. The time (h:min:s:ms) is indicated (top right). (MOV 3353 kb)

In metaphase I, the chromosome shape is not altered by TEV protease in ΔNcaph1Δ Ncaph2 oocytes rescued by wild type Ncaph2.

Oocytes from Ncaph1f/f Ncaph2f/f Tg(ZP3Cre) females were isolated in M16 supplemented with IBMX and injected at GV stage by mRNA encoding mCherry tagged H2B histone, TALE-mClover_MajSat, WT-NCAPH2 and MAD2. After two hours of incubation to allow the expression of the injected mRNA, IBXM was washed out and oocytes incubated overnight. Metaphase I arrested oocytes were then injected for a second time, with mRNA encoding TEV protease and followed by live cell confocal microscopy. The time (h:min:s:ms) is indicated (top right). (MOV 3547 kb)

In metaphase I, TEV protease induces the rapid stretching of chromosomes in ΔNcaph1ΔNcaph2 oocytes rescued by TEV-Ncaph2.

Oocytes from Ncaph1f/f Ncaph2f/f Tg(ZP3Cre) females were isolated in M16 supplemented with IBMX and injected at GV stage by mRNA encoding mCherry tagged H2B histone, TALE-mClover_MajSat, TEV-NCAPH2 and MAD2. After two hours of incubation to allow the expression of the injected mRNA, IBXM was washed out and oocytes incubated overnight. Metaphase I arrested oocytes were then injected for a second time, with mRNA encoding TEV protease and followed by live cell confocal microscopy. The time (h:min:s:ms) is indicated (top right). (MOV 4862 kb)

In metaphase II, the chromosome morphology is not altered by TEV protease in ΔNcaph2 oocytes rescued by wild type Ncaph2.

Oocytes from Ncaph2f/f Tg(ZP3Cre) females were isolated in M16 supplemented with IBMX and injected at GV stage by mRNA encoding mCherry tagged H2B histone, TALE-mClover_MajSat and WT-NCAPH2. After two hours of incubation to allow the expression of the injected mRNA, IBXM was washed out and oocytes incubated overnight. Metaphase II arrested oocytes were then injected for a second time, with mRNA encoding TEV protease and followed by live cell confocal microscopy. The time (h:min:s:ms) is indicated (top right). (MOV 2962 kb)

In metaphase II, the chromosome morphology is not altered by TEV protease in ΔNcaph2 oocytes rescued by TEV-Ncaph2.

Oocytes from Ncaph2f/f Tg(ZP3Cre) females were isolated in M16 supplemented with IBMX and injected at GV stage by mRNA encoding mCherry tagged H2B histone, TALE-mClover_MajSat and TEV-NCAPH2. After two hours of incubation to allow the expression of the injected mRNA, IBXM was washed out and oocytes incubated overnight. Metaphase II arrested oocytes were then injected for a second time, with mRNA encoding TEV protease and followed by live cell confocal microscopy. The time (h:min:s:ms) is indicated (top right). (MOV 2064 kb)

In metaphase I, the co-injection of wild type Ncaph2 does not rescue the unravelling of the chromosomes induced by the TEV protease.

Oocytes from Ncaph2f/f Tg(ZP3Cre) females were isolated in M16 supplemented with IBMX and injected at GV stage by mRNA encoding mCherry tagged H2B histone, TALE-mClover_MajSat, TEV-NCAPH2 and MAD2. After two hours of incubation to allow the expression of the injected mRNA, IBXM was washed out and oocytes incubated overnight. Metaphase I arrested oocytes were then injected for a second time, with mRNA encoding TEV protease and WT-NCAPH2 and followed by live cell confocal microscopy. The time (h:min:s:ms) is indicated (top right). (MOV 3021 kb)

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Houlard, M., Godwin, J., Metson, J. et al. Condensin confers the longitudinal rigidity of chromosomes. Nat Cell Biol 17, 771–781 (2015). https://doi.org/10.1038/ncb3167

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/ncb3167

This article is cited by

Search

Advanced 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