Studying G2 DNA Damage Checkpoints Using the Fission Yeast Schizosaccharomyces Pombe

Overview

Journal Methods Mol Biol

Specialty Molecular Biology

Date 2011 Aug 27

PMID 21870280

Citations 6

Authors

Nicholas Willis

Nicholas Rhind

Affiliations

Soon will be listed here.

Abstract

Using synchronized cells, one can directly measure delay in mitosis brought about by the G2 DNA damage checkpoint in response to exposure to exogenous DNA damaging agents. Scoring mitosis in the fission yeast Schizosaccharomyces pombe is relatively simple. Many techniques exist for synchronizing cells for such assays. We present a detailed explanation of the setup and use of centrifugal elutriation to synchronize cells in G2, exposure of cells to DNA damage, and measurement of mitotic progression and delay.

Citing Articles

Cell Cycle Progression and Synchronization: An Overview.

Wang Z Methods Mol Biol. 2022; 2579:3-23.

PMID: 36045194 DOI: 10.1007/978-1-0716-2736-5_1.

An Optimized and Versatile Counter-Flow Centrifugal Elutriation Workflow to Obtain Synchronized Eukaryotic Cells.

Liu Y, Nan B, Niu J, Kapler G, Gao S Front Cell Dev Biol. 2021; 9:664418.

PMID: 33959616 PMC: 8093812. DOI: 10.3389/fcell.2021.664418.

The fission yeast S-phase cyclin Cig2 can drive mitosis.

Pickering M, Magner M, Keifenheim D, Rhind N Genetics. 2021; 217(1):1-12.

PMID: 33683349 PMC: 8045716. DOI: 10.1093/genetics/iyaa002.

Gene Expression of Pneumocystis murina after Treatment with Anidulafungin Results in Strong Signals for Sexual Reproduction, Cell Wall Integrity, and Cell Cycle Arrest, Indicating a Requirement for Ascus Formation for Proliferation.

Cushion M, Ashbaugh A, Hendrix K, Linke M, Tisdale N, Sayson S Antimicrob Agents Chemother. 2018; 62(5).

PMID: 29463544 PMC: 5923105. DOI: 10.1128/AAC.02513-17.

Size-Dependent Expression of the Mitotic Activator Cdc25 Suggests a Mechanism of Size Control in Fission Yeast.

Keifenheim D, Sun X, DSouza E, Ohira M, Magner M, Mayhew M Curr Biol. 2017; 27(10):1491-1497.e4.

PMID: 28479325 PMC: 5479637. DOI: 10.1016/j.cub.2017.04.016.

References

Sullivan D, Huffaker T . Astral microtubules are not required for anaphase B in Saccharomyces cerevisiae. J Cell Biol. 1992; 119(2):379-88. PMC: 2289657. DOI: 10.1083/jcb.119.2.379. View

Edwards R, Carr A . Analysis of radiation-sensitive mutants of fission yeast. Methods Enzymol. 1997; 283:471-94. DOI: 10.1016/s0076-6879(97)83038-8. View

Stewart E, Enoch T . S-phase and DNA-damage checkpoints: a tale of two yeasts. Curr Opin Cell Biol. 1996; 8(6):781-7. DOI: 10.1016/s0955-0674(96)80078-0. View

Rhind N, Russell P . Mitotic DNA damage and replication checkpoints in yeast. Curr Opin Cell Biol. 1999; 10(6):749-58. PMC: 2864141. DOI: 10.1016/s0955-0674(98)80118-x. View

Sancar A, Lindsey-Boltz L, Unsal-Kacmaz K, Linn S . Molecular mechanisms of mammalian DNA repair and the DNA damage checkpoints. Annu Rev Biochem. 2004; 73:39-85. DOI: 10.1146/annurev.biochem.73.011303.073723. View