This proposal aims at further understanding the molecular composition, the higher-order structure and the biological function of the budding yeast kinetochore have been cloned and several protein components have been identified. However, much less is known about its higher-order structure and the mechanisms of its functions. In this proposal, two in vitro assays have been designed: 1. By a cross-linking/PCR, specific protein/DNA interaction can be detected. This assay will be applied to the study of the kinetochore and the cell cycle to study the role of the DNA and protein elements in the assembly of the kinetochore and the cell cycle regulation of the kinetochore assembly, to define its higher-order structure and to confirm the localization of putative kinetochore proteins. 2. By the combination of the yeast chromosome tagging, minichromosome isolation and an in vitro chromosome/microtubule interaction assay techniques, the biochemical activities of the yeast kinetochore on a minichromosome can me measured in vitro. This assay can be applied to the study of the kinetochore functions, specifically, the kinetics of the kinetochore functions, specifically, the kinetics of the kinetochore/microtubule interaction, the cell cycle regulation of its activities and the contribution of specific DNA and protein elements to the kinetochore activities.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Postdoctoral Individual National Research Service Award (F32)
Project #
5F32GM019483-03
Application #
6179865
Study Section
Biological Sciences 2 (BIOL)
Program Officer
Tompkins, Laurie
Project Start
1998-04-15
Project End
Budget Start
2000-04-15
Budget End
2001-04-14
Support Year
3
Fiscal Year
2000
Total Cost
$37,516
Indirect Cost
Name
Massachusetts Institute of Technology
Department
Biology
Type
Schools of Arts and Sciences
DUNS #
City
Cambridge
State
MA
Country
United States
Zip Code
02139