The proper chromosome segregation during cell division depends on the maintenance and regulation of the association between sister chromatids. To attain a clear understanding of sister-chromatid cohesion a study of the protein. MEI-S332, will be implemented. The objectives of the experiments in this proposal will determine where MEI-S332 binds to on the centromeric region of meiotic chromosomes and to determine how its disappearance at the metaphase II/anaphase II transition is regulated. The fulfillment of these objectives is essential for understanding the regulation of sister- chromatid cohesion. The following questions regarding MEI-S332's function will be tested: 1) Is MEI-S332 bound to the heterochromatin flanking the centromere or to the kinetochore itself? The localization of MEI-S332-GFP on an inverted chromosome and on deleted minichromosomes will be used for the purpose. 2) Is MEI-S332's disappearance from the centromeric region at anaphase II a result of cell cycle regulated proteolysis or post translational modification such as phosphorylation? Western blot analysis will be performed to test which of these mechanisms regulate MEI-S332 localization. 3) What regions of the protein dictate its localization to the centromeric region? A site-directed mutagenesis will be utilized to define essential regions of MEI-S332 that dictate its localization and disappearance. 4) What effects do other mutations have on MEI-S332-GFP behavior will be examined in the genetic background of mutants that are defective in chromosome segregation and cell cycle control during meiosis.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Postdoctoral Individual National Research Service Award (F32)
Project #
5F32GM018433-02
Application #
2637922
Study Section
Special Emphasis Panel (ZRG2-GEN (05))
Project Start
1998-01-01
Project End
Budget Start
1998-01-01
Budget End
1998-12-31
Support Year
2
Fiscal Year
1998
Total Cost
Indirect Cost
Name
Whitehead Institute for Biomedical Research
Department
Type
DUNS #
076580745
City
Cambridge
State
MA
Country
United States
Zip Code
02142