E-Locus - Institutional Repository of the University of Crete - Ταυτοποίηση νέων πρωτεϊνών που αλληλεπιδρούν με την Chk1 κατά την ενεργοποίηση του spindle checkpoint

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Identifier

000363348

Title

Ταυτοποίηση νέων πρωτεϊνών που αλληλεπιδρούν με την Chk1 κατά την ενεργοποίηση του spindle checkpoint

Alternative Title

Identification of novel Chk1 interacting proteins in the spindle checkpoint

Author

Περάκη, Ιωάννα

Thesis advisor

Ζάχος, Γιώργος
Τζαμαρίας, Δημήτρης
Χαλεπάκης, Γιώργος

Abstract

Τhe spindle checkpoint blocks initiation of anaphase when chromosomes are not bipolarly attached to spindle microtubules and protects against chromosome mis-segregation. Τhe spindle checkpoint is triggered by unattached kinetochores or lack of tension across attached sister kinetochores. Chk1 has well-defined roles in the DNA damage checkpoints, but the role and substrates of Chk1 in the spindle checkpoint have not been fully described. We used two approaches to identify novel proteins that interact with Chk1 during activation of the spindle checkpoint by taxol in vertebrate cells : a) Using chemical genetics, we investigated whether avian DT40 Chk1^as cells expressing a mutant Chk1 which is sensitive to inhibition by bulky compounds NM-PP1 and 3MB-PP1, may be used to identify novel Chk1 substrates in the G2/M DNA damage and spindle checkpoints, b) We isolated mitotic spindles extracts from human HeLa cells and detected Chk1-interacting proteins by immunoprecipitation experiments. We showed that: a) DT40 Chk1^as cells might be used to identify Chk1 substrates in the G2/M DNA damage, but not the spindle checkpoint, b) We detected Chk1-interacting candidate proteins with approximate masses of 160, 105 and 75 kDa and will identify them by mass-spectrometry. Identifying novel Chk1 interacting proteins in the spindle checkpoint will help us to better understand the mechanisms of mitotic cell division.

Language

Greek

Subject

Chemical genetics

Protein interactions

Substrates

Πρωτεϊνικές αλληλεπιδράσεις

Υποστρώματα

Χημική γενετική

Issue date

2011-03-18

Collection