Molecular architecture of the Saccharomyces cerevisiae kinetochore
Thesis (Ph. D.)--Massachusetts Institute of Technology, Dept. of Biology, 2004.
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| Format: | Thesis |
| Language: | en_US |
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Massachusetts Institute of Technology
2005
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| Online Access: | http://hdl.handle.net/1721.1/28680 |
| _version_ | 1826211562525818880 |
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| author | Espelin, Christopher W., 1969- |
| author2 | Peter Sorger. |
| author_facet | Peter Sorger. Espelin, Christopher W., 1969- |
| author_sort | Espelin, Christopher W., 1969- |
| collection | MIT |
| description | Thesis (Ph. D.)--Massachusetts Institute of Technology, Dept. of Biology, 2004. |
| first_indexed | 2024-09-23T15:07:58Z |
| format | Thesis |
| id | mit-1721.1/28680 |
| institution | Massachusetts Institute of Technology |
| language | en_US |
| last_indexed | 2024-09-23T15:07:58Z |
| publishDate | 2005 |
| publisher | Massachusetts Institute of Technology |
| record_format | dspace |
| spelling | mit-1721.1/286802019-04-11T11:58:26Z Molecular architecture of the Saccharomyces cerevisiae kinetochore Molecular architecture of the S. cerevisiae kinetochore Espelin, Christopher W., 1969- Peter Sorger. Massachusetts Institute of Technology. Dept. of Biology. Massachusetts Institute of Technology. Dept. of Biology. Biology. Thesis (Ph. D.)--Massachusetts Institute of Technology, Dept. of Biology, 2004. Includes bibliographical references. (cont.) checkpoint proteins Bub lp and Bub3p are recruited to the kinetochore as part of the normal cell cycle in budding yeast, whereas Madlp and Mad2p are recruited only in response to damage to the CEN-kinetochore-MT connection. Analysis of specific S. cerevisiae kinetochore mutants indicates that attachment status is the signal recognized by the spindle checkpoint to allow the metaphase-to-anaphase transition to occur. The kinetochore is a multiprotein complex capable of simultaneously binding centromeric DNA (CEN) and microtubules (MTs). In this capacity, the kinetochore is responsible for enabling sister chromatids to become attached to the mitotic spindle, thus ensuring proper chromosome segregation. Due to its small size and simple structure, the Saccharomyces cerevisiae CEN is particularly well suited for studying kinetochore assembly and function. Analysis of S. cerevisiae Ndc lop established the binding of this kinetochore protein to the CDEII element of the CEN, forming part of the DNA-Binding layer upon which the rest of the kinetochore is assembled. Alteration of the nucleotide bases deemed important for Ndc lOp-CEN binding il vitro, have dramatic effects on chromosome segregation in vivo. Further characterization indicates that Ndc lop may also play a role in chromosome segregation at non-CEN locations. A combination of live-cell imaging, biochemical and genetic techniques were used to identify eleven novel S. cerevisiae kinetochore subunits and elucidate their roles in microtubule attachment. Chromatin Immunoprecipitation (ChIP) Assays evaluated both the range of CEN DNA bound by these kinetochore proteins and determined the interdependencies required for their association with CEN DNA. Using this data, a preliminary model of the molecular architecture of the kinetochore is beginning to emerge. Dynabead-Kinetochore Reconstitution Assays suggest an interaction between the CBF3 Complex bound to CEN DNA and Ndc80p, Cse4p, Mif2p and Amelp, adding further insight into kinetochore assembly. The spindle assembly checkpoint monitors proper bipolar attachment of sister chromatids to the mitotic spindle. Microscopy and ChIP demonstrate by Christopher W. Espelin. Ph.D. 2005-09-27T17:41:50Z 2005-09-27T17:41:50Z 2004 2004 Thesis http://hdl.handle.net/1721.1/28680 58995239 en_US M.I.T. theses are protected by copyright. They may be viewed from this source for any purpose, but reproduction or distribution in any format is prohibited without written permission. See provided URL for inquiries about permission. http://dspace.mit.edu/handle/1721.1/7582 242 leaves 15224583 bytes 15256466 bytes application/pdf application/pdf application/pdf Massachusetts Institute of Technology |
| spellingShingle | Biology. Espelin, Christopher W., 1969- Molecular architecture of the Saccharomyces cerevisiae kinetochore |
| title | Molecular architecture of the Saccharomyces cerevisiae kinetochore |
| title_full | Molecular architecture of the Saccharomyces cerevisiae kinetochore |
| title_fullStr | Molecular architecture of the Saccharomyces cerevisiae kinetochore |
| title_full_unstemmed | Molecular architecture of the Saccharomyces cerevisiae kinetochore |
| title_short | Molecular architecture of the Saccharomyces cerevisiae kinetochore |
| title_sort | molecular architecture of the saccharomyces cerevisiae kinetochore |
| topic | Biology. |
| url | http://hdl.handle.net/1721.1/28680 |
| work_keys_str_mv | AT espelinchristopherw1969 moleculararchitectureofthesaccharomycescerevisiaekinetochore AT espelinchristopherw1969 moleculararchitectureofthescerevisiaekinetochore |