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DC Field | Value | Language |
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dc.contributor.author | Ribeiro, SA | - |
dc.contributor.author | Vagnarelli, P | - |
dc.contributor.author | Dong, Y | - |
dc.contributor.author | Hori, T | - |
dc.contributor.author | McEwen, BF | - |
dc.contributor.author | Fukagawa, T | - |
dc.contributor.author | Flors, C | - |
dc.contributor.author | Earnshaw, WC | - |
dc.date.accessioned | 2014-08-20T09:15:58Z | - |
dc.date.available | 2014-08-20T09:15:58Z | - |
dc.date.issued | 2010 | - |
dc.identifier.citation | Proceedings of the National Academy of Sciences, 107(23), 10484 - 10489, 2010 | en_US |
dc.identifier.issn | 1091-6490 | - |
dc.identifier.uri | http://www.pnas.org/content/107/23/10484 | en |
dc.identifier.uri | http://bura.brunel.ac.uk/handle/2438/8940 | - |
dc.description | This article is freely available online through the PNAS open access option. Copyright @ 2010 National Academy of Sciences. | en_US |
dc.description.abstract | A longstanding question in centromere biology has been the organization of CENP-A–containing chromatin and its implications for kinetochore assembly. Here, we have combined genetic manipulations with deconvolution and super-resolution fluorescence microscopy for a detailed structural analysis of chicken kinetochores. Using fluorescence microscopy with subdiffraction spatial resolution and single molecule sensitivity to map protein localization in kinetochore chromatin unfolded by exposure to a low salt buffer, we observed robust amounts of H3K9me3, but only low levels of H3K4me2, between CENP-A subdomains in unfolded interphase prekinetochores. Constitutive centromere-associated network proteins CENP-C and CENP-H localize within CENP-A–rich subdomains (presumably on H3-containing nucleosomes) whereas CENP-T localizes in interspersed H3-rich blocks. Although interphase prekinetochores are relatively more resistant to unfolding than sur-rounding pericentromeric heterochromatin, mitotic kinetochores are significantly more stable, reflecting mitotic kinetochore maturation. Loss of CENP-H, CENP-N, or CENP-W had little or no effect on the unfolding of mitotic kinetochores. However, loss of CENP-C caused mitotic kinetochores to unfold to the same extent as their interphase counterparts. Based on our results we propose a new model for inner centromeric chromatin architecture in which chromatin is folded as a layered boustrophedon, with planar sinusoids containing interspersed CENP-A–rich and H3-rich subdomains oriented toward the outer kinetochore. In mitosis, a CENP-C–dependent mechanism crosslinks CENP-A blocks of different layers together, conferring extra stability to the kinetochore. | en_US |
dc.description.sponsorship | The Wellcome Trust, the EPSRC, the Darwin Trust of Edinburgh, the NIH, and the Ministry of Education, Culture, Sports, Science and Technology of Japan. | en_US |
dc.language | eng | - |
dc.language.iso | en | en_US |
dc.publisher | National Academy of Sciences | en_US |
dc.subject | CENP-A | en_US |
dc.subject | Centromere | en_US |
dc.subject | Super-resolution imaging | en_US |
dc.subject | Chromosome | en_US |
dc.subject | Boustrophedon | en_US |
dc.title | A super-resolution map of the vertebrate kinetochore | en_US |
dc.type | Article | en_US |
dc.identifier.doi | http://dx.doi.org/10.1073/pnas.1002325107 | - |
pubs.organisational-data | /Brunel | - |
pubs.organisational-data | /Brunel/Brunel Staff by College/Department/Division | - |
pubs.organisational-data | /Brunel/Brunel Staff by College/Department/Division/College of Health and Life Sciences | - |
pubs.organisational-data | /Brunel/Brunel Staff by College/Department/Division/College of Health and Life Sciences/Dept of Life Sciences | - |
pubs.organisational-data | /Brunel/Brunel Staff by College/Department/Division/College of Health and Life Sciences/Dept of Life Sciences/Biological Sciences | - |
pubs.organisational-data | /Brunel/University Research Centres and Groups | - |
pubs.organisational-data | /Brunel/University Research Centres and Groups/School of Health Sciences and Social Care - URCs and Groups | - |
pubs.organisational-data | /Brunel/University Research Centres and Groups/School of Health Sciences and Social Care - URCs and Groups/Brunel Institute for Ageing Studies | - |
Appears in Collections: | Biological Sciences Dept of Life Sciences Research Papers |
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Fulltext.pdf | 8.55 MB | Adobe PDF | View/Open |
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