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Identification and characterization ...
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Page, Andrew M.
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Identification and characterization of new subunits of the Saccharomyces cerevisiae anaphase promoting complex.
紀錄類型:
書目-電子資源 : Monograph/item
正題名/作者:
Identification and characterization of new subunits of the Saccharomyces cerevisiae anaphase promoting complex./
作者:
Page, Andrew M.
面頁冊數:
207 p.
附註:
Source: Dissertation Abstracts International, Volume: 65-12, Section: B, page: 6200.
Contained By:
Dissertation Abstracts International65-12B.
標題:
Biology, Molecular. -
電子資源:
http://pqdd.sinica.edu.tw/twdaoapp/servlet/advanced?query=3155664
ISBN:
9780496164684
Identification and characterization of new subunits of the Saccharomyces cerevisiae anaphase promoting complex.
Page, Andrew M.
Identification and characterization of new subunits of the Saccharomyces cerevisiae anaphase promoting complex.
- 207 p.
Source: Dissertation Abstracts International, Volume: 65-12, Section: B, page: 6200.
Thesis (Ph.D.)--The Johns Hopkins University, 2005.
Ubiquitin-mediated proteolysis is an essential component of cell cycle regulation. The Anaphase-Promoting Complex/Cyclosome (APC/C) is a large, multisubunit E3 ubiquitin ligase required for the ubiquitination of substrates whose proteolysis drives the metaphase to anaphase transition and exit from mitosis. Substrates which are polyubiquitinated by the APC/C are subsequently degraded by the proteasome. Destruction of these substrates, which include securin (Pds1p) and the mitotic cyclins (Clb1-6p), is essential for the successful completion of chromosome segregation and cytokinesis. The APC/C is conserved throughout the eukaryotic kingdom and comprises at least a dozen subunits. In this work we have identified four new APC/C subunits in the budding yeast Saccharomyces cerevisiae: Apc2p, Apc11p, Swm1p, and Mnd2p. Two subunits, Apc2p and Apc11p, are encoded by essential genes, while deletion of Mnd2p and Swm1p is not deleterious to cell viability at permissive temperatures. We have demonstrated both physical and genetic interactions of these new components with previously identified APC/C subunits. Furthermore, we have characterized the requirement of each of these subunits, along with that of another subunit, Apc9p, for APC/C function in vivo. Apc2p, Apc11p, and Swm1p are required for the efficient proteolysis of multiple APC/C substrates. For strains lacking Swm1p, we observed significant APC/C substrate proteolysis defects even at temperatures permissive for cell viability. In the case of Apc9p, we observed some delay in the onset of Clb2p degradation, but other APC/C substrates, including Pds1p and Clb5p, were degraded with wild-type kinetics. In contrast, strains lacking Mnd2p did not exhibit any substantial defects in APC/C-mediated proteolysis. Our results demonstrate that different APC/C subunits make distinct contributions to APC/C function and suggest that certain non-essential APC/C subunits are required for the degradation of some substrates, but not others. Finally, we have begun to investigate the localization of APC/C subunits. We have observed APC/C subunit binding to chromatin in chromosome spreads and localizing as punctate foci in live or lightly fixed cells. Our preliminary results suggest that the APC/C is sequestered in punctate foci when the APC/C is inactive and disperses throughout the nucleus upon APC/C activation from anaphase onset to the end of G1.
ISBN: 9780496164684Subjects--Topical Terms:
1017719
Biology, Molecular.
Identification and characterization of new subunits of the Saccharomyces cerevisiae anaphase promoting complex.
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Ubiquitin-mediated proteolysis is an essential component of cell cycle regulation. The Anaphase-Promoting Complex/Cyclosome (APC/C) is a large, multisubunit E3 ubiquitin ligase required for the ubiquitination of substrates whose proteolysis drives the metaphase to anaphase transition and exit from mitosis. Substrates which are polyubiquitinated by the APC/C are subsequently degraded by the proteasome. Destruction of these substrates, which include securin (Pds1p) and the mitotic cyclins (Clb1-6p), is essential for the successful completion of chromosome segregation and cytokinesis. The APC/C is conserved throughout the eukaryotic kingdom and comprises at least a dozen subunits. In this work we have identified four new APC/C subunits in the budding yeast Saccharomyces cerevisiae: Apc2p, Apc11p, Swm1p, and Mnd2p. Two subunits, Apc2p and Apc11p, are encoded by essential genes, while deletion of Mnd2p and Swm1p is not deleterious to cell viability at permissive temperatures. We have demonstrated both physical and genetic interactions of these new components with previously identified APC/C subunits. Furthermore, we have characterized the requirement of each of these subunits, along with that of another subunit, Apc9p, for APC/C function in vivo. Apc2p, Apc11p, and Swm1p are required for the efficient proteolysis of multiple APC/C substrates. For strains lacking Swm1p, we observed significant APC/C substrate proteolysis defects even at temperatures permissive for cell viability. In the case of Apc9p, we observed some delay in the onset of Clb2p degradation, but other APC/C substrates, including Pds1p and Clb5p, were degraded with wild-type kinetics. In contrast, strains lacking Mnd2p did not exhibit any substantial defects in APC/C-mediated proteolysis. Our results demonstrate that different APC/C subunits make distinct contributions to APC/C function and suggest that certain non-essential APC/C subunits are required for the degradation of some substrates, but not others. Finally, we have begun to investigate the localization of APC/C subunits. We have observed APC/C subunit binding to chromatin in chromosome spreads and localizing as punctate foci in live or lightly fixed cells. Our preliminary results suggest that the APC/C is sequestered in punctate foci when the APC/C is inactive and disperses throughout the nucleus upon APC/C activation from anaphase onset to the end of G1.
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