A Method for Analyzing the Ubiquitination and Degradation

A technical article on method for analyzing the ubiquitination and degradation from Biological Procedures Online.

Biol. Proced. Online 2002;4:62-69

A Method for Analyzing the Ubiquitination and Degradation of Aurora-A

Andrea Klotzbucher, Gaetan Pascreau, Claude Prigent and Yannick Arlot-Bonnemains

Abstract:

The cell cycle machinery consists of regulatory proteins that control the progression through the cell cycle ensuring that DNA replication alternates with DNA segregation in mitosis to maintain cell integrity. Some of these key regulators have to be degraded at each cell cycle to prevent cellular dysfunction. Mitotic exit requires the inactivation of cyclin dependent kinase1 (cdk1) and it is the degradation of the cyclin subunit that inactivates the kinase. Cyclin degradation has been well characterized and it was shown that it is ubiquitin proteasome pathway that leads to the elimination of cyclins. By now, many other regulatory proteins were shown to be degraded by the same pathway, among them members of the aurora kinase family, degraded many other regulatory proteins. Aurora kinases are involved in mitotic spindle formation as well as in cytokinesis. The abundance and activity of the kinase is precisely regulated during the cell cycle. To understand how proteolysis regulates transitions through the cell cycle we describe two assays for ubiquitination and degradation of xenopus aurora kinase A using extracts from xenopus eggs or somatic cell lines.

Archived page

Last update 20-Oct-2005, Rating n/a of 0 votes.