The flawless execution of cell division is essential to the generation and survival of all organisms. During every cell cycle, chromosomes must be accurately partitioned to daughter cells to prevent genomic instability and aneuploidy, a hallmark of many tumors and birth defects. Chromosomes segregate using their kinetochores, the specialized protein structures that assemble on centromeric DNA sequences and mediate attachment to microtubules. The foundation of all eukaryotic kinetochores is a conserved, inner centromere structure characterized by centromeric chromatin and its associated proteins. A hallmark of centromeric chromatin is Cenp-A, an essential histone H3 variant that epigenetically marks centromeres and is required for kinetochore assembly. The surrounding pericentromeric chromatin also makes various contributions to the fidelity of segregation. To fully understand the mechanisms that ensure accurate chromosome segregation, it is critical to elucidate outstanding questions about the functions and maintenance of centromeric and pericentromeric chromatin. This proposal will use purified kinetochores and a combination of biochemical, biophysical, proteomic and genomic approaches to address a number of outstanding questions about the contribution of centromeric chromatin to kinetochore function. 1) How do centromere-binding proteins contribute to the diverse functions of kinetochores? 2) How does pericentromeric chromatin regulate chromosome segregation? 3) What are the mechanisms that contribute to the exclusive localization of centromeres? The proposal will use budding yeast for these studies because they are amenable to biochemical, genetic and cytological studies, and the yeast kinetochore is the best characterized to date. Taken together, these studies of kinetochores and the underlying chromatin foundation in budding yeast will lead toward an understanding of the fundamental mechanisms of segregation in all eukaryotes. This work will not only elucidate important aspects about the process of segregation, but will aid in the design of better therapeutic interventions in the long-term.

Public Health Relevance

All cells must inherit the right number of chromosomes every time they divide because the wrong number of chromosomes is a hallmark of cancer, birth defects, and other diseases related to problems in cell proliferation. We are therefore studying the process of chromosome partitioning to daughter cells when they divide to understand the basis for a number of human diseases.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM078069-06
Application #
8423684
Study Section
Cellular Signaling and Regulatory Systems Study Section (CSRS)
Program Officer
Carter, Anthony D
Project Start
2007-09-15
Project End
2016-01-31
Budget Start
2013-02-01
Budget End
2014-01-31
Support Year
6
Fiscal Year
2013
Total Cost
$362,712
Indirect Cost
$150,412
Name
Fred Hutchinson Cancer Research Center
Department
Type
DUNS #
078200995
City
Seattle
State
WA
Country
United States
Zip Code
98109
Hildebrand, Erica M; Biggins, Sue (2016) Regulation of Budding Yeast CENP-A levels Prevents Misincorporation at Promoter Nucleosomes and Transcriptional Defects. PLoS Genet 12:e1005930
London, Nitobe; Biggins, Sue (2014) Signalling dynamics in the spindle checkpoint response. Nat Rev Mol Cell Biol 15:736-47
Driver, Jonathan W; Powers, Andrew F; Sarangapani, Krishna K et al. (2014) Measuring kinetochore-microtubule interaction in vitro. Methods Enzymol 540:321-37
Biggins, Sue; Welch, Matthew D (2014) Editorial overview: Cell architecture: Cellular organization and function. Curr Opin Cell Biol 26:v-vii
Deyter, Gary M R; Biggins, Sue (2014) The FACT complex interacts with the E3 ubiquitin ligase Psh1 to prevent ectopic localization of CENP-A. Genes Dev 28:1815-26
Biggins, Sue (2013) The composition, functions, and regulation of the budding yeast kinetochore. Genetics 194:817-46
Akiyoshi, Bungo; Nelson, Christian R; Biggins, Sue (2013) The aurora B kinase promotes inner and outer kinetochore interactions in budding yeast. Genetics 194:785-9
Ng, Tessie M; Lenstra, Tineke L; Duggan, Nicole et al. (2013) Kinetochore function and chromosome segregation rely on critical residues in histones H3 and H4 in budding yeast. Genetics 195:795-807
Akiyoshi, Bungo; Nelson, Christian R; Duggan, Nicole et al. (2013) The Mub1/Ubr2 ubiquitin ligase complex regulates the conserved Dsn1 kinetochore protein. PLoS Genet 9:e1003216
Gonen, Shane; Akiyoshi, Bungo; Iadanza, Matthew G et al. (2012) The structure of purified kinetochores reveals multiple microtubule-attachment sites. Nat Struct Mol Biol 19:925-9

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