The Mitotic Exit Network: Methods and Protocols

Part I: Analysis of Cell Cycle Progression and Mitotic Exit

1. Regulation of Mitotic Exit in Saccharomyces cerevisiae

            Bàrbara Baro, Ethel Queralt, and Fernando Monje-Casas

2. Methods of Synchronization of Yeast Cells for the Analysis of Cell Cycle Progression

            M. Angeles Juanes

3. Analysis of Cell Cycle Progression in Saccharomyces cerevisiae Using the Budding Index and Tubulin Staining

4. Determination of Cell Cycle Stage and Mitotic Exit Through the Quantification of the Protein Levels of Known Mitotic Regulators

            Cristina Cepeda-García

5. Cdc14 Localization as a Marker for Mitotic Exit: In Vivo Quantitative Analysis of Cdc14 Release

            Gabriel Neurohr and Manuel Mendoza

Part II: Biochemistry of the MEN

            Marco Geymonat, Adonis Spanos, and Katrin Rittinger

7. Analysis of Protein-Protein Interactions Between MEN Components

            Rami Rahal

8. A New Methodology for the Quantification of In Vivo Cdc14 Phosphatase Activity

            Ethel Queralt and José-Antonio Rodriguez-Rodriguez

9. Analysis of SUMOylation in the RENT Complex by Fusion to a SUMO-Specific Protease Domain

            Neus Colomina, Clàudia Guasch, and Jordi Torres-Rosell

10. A Substrate Trapping Method for Identification of Direct Cdc14 Phosphatase Targets

            Brendan L. Powers, Hana Hall, Harry Charbonneau, and Mark C. Hall

Part III: Analysis of the Localization of MEN Components

11. Localizing MEN Components by Indirect Immunofluorescence Analysis of Budding Yeast

12. Analysis of the Localization of MEN Components by Live Cell Imaging Microscopy

            Zhiang Guo and Marisa Segal

13. Evaluation of the Dynamicity of Mitotic Exit Network and Spindle Position Checkpoint Components on Spindle Pole Bodies by Fluorescence Recovery After Photobleaching (FRAP)

            Ayse Koca Caydasi and Gislene Pereira

14. Asymmetric Localization of Components and Regulators of the Mitotic Exit Network at Spindle Pole Bodies

            Ilaria Scarfone and Simonetta Piatti

15. Evaluation of the Nucleolar Localization of the RENT Complex to Ribosomal DNA by Chromatin Immunoprecipitation Assays

            Julie Huang, Nahid Iglesias, and Danesh Moazed

Part IV: Control of Cellular Processes by the MEN

16. Analysis of the Functionality of the Mitotic Checkpoints

            Roberta Fraschini

            Facundo Ramos, Joanne Leonard, Andrés Clemente-Blanco, and Luis Aragón

18. Studying the Role of the Mitotic Exit Network in Cytokinesis

            Magdalena Foltman and Alberto Sanchez-Diaz

Part V: Final Remarks: The MEN as a Model for Higher Eukaryotes

19. Hippo Signaling in Mitosis: An Updated View in Light of the MEN Pathway