Sister chromatid cohesion is an essential cellular process that ensures the faithful segregation of genetic information during mitosis and meiosis. The long-term goal of this project is to understand the biochemical basis of this process by using cell-free extracts derived from Xenopus laevis (African toad) eggs and mammalian tissue culture cells. An emphasis will be made on the structural and functional characterization of cohesin, a highly conserved protein complex that plays a key role in establishing and maintaining sister chromatid cohesion. The cohesin complex is composed of two SMC (structural maintenance of chromosomes) subunits and at least two non-S MC subunits, and is predicted to act as part of the molecular """"""""glue"""""""" that holds two sister chromatids together. In this proposal, (1) Biochemical and structural approaches will be combined to understand the molecular mechanisms of action of the cohesin complex. (2) Cell cycle-dependent interactions between cohesin and chromatin will be reconstituted in vitro and the molecular basis of loading and unloading processes will be determined. (3) Vertebrate homologs of BimD and Scc2, two proteins implicated genetically in sister chromatid cohesion, will be identified and characterized in the cell-free extracts and in tissue culture cells. (4) A novel in vitro assay will be developed to determine how sister chromatid cohesion is functionally coupled to DNA replication. The information obtained from this work will ultimately contribute to a better understanding of human health because chromosome anomalies, such as aneuploidy and translocations, are tightly associated with tumor development and birth defects.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
1R01GM063545-01
Application #
6359257
Study Section
Cell Development and Function Integrated Review Group (CDF)
Program Officer
Carter, Anthony D
Project Start
2001-07-01
Project End
2005-06-30
Budget Start
2001-07-01
Budget End
2002-06-30
Support Year
1
Fiscal Year
2001
Total Cost
$292,355
Indirect Cost
Name
Cold Spring Harbor Laboratory
Department
Type
DUNS #
065968786
City
Cold Spring Harbor
State
NY
Country
United States
Zip Code
11724
Gandhi, Rita; Gillespie, Peter J; Hirano, Tatsuya (2006) Human Wapl is a cohesin-binding protein that promotes sister-chromatid resolution in mitotic prophase. Curr Biol 16:2406-17
Losada, Ana; Yokochi, Tomoki; Hirano, Tatsuya (2005) Functional contribution of Pds5 to cohesin-mediated cohesion in human cells and Xenopus egg extracts. J Cell Sci 118:2133-41