Study of the DNA damage checkpoint using Xenopus egg extracts

Journal of Visualized Experiments

Volumn , Issue 69, 2012, Pages

  Willis, Jeremy ^a   Destephanis, Darla ^a   Patel, Yogin ^a   Gowda, Vrushab ^a   Yan, Shan ^a  

^a UNIVERSITY OF NORTH CAROLINA AT CHARLOTTE   (United States)

Author keywords

AT70; ATR; Cellular biology; Chk1 phosphorylation; Developmental biology; DNA damage checkpoint; Genetics; Immunoblotting; Issue 69; MMS; Molecular biology; UV; Xenopus egg extracts; Xenopus laevis

Indexed keywords

ATR PROTEIN, XENOPUS; CELL CYCLE PROTEIN; CHECKPOINT KINASE 1; DNA; MITOMYCIN; PROTEIN KINASE; PROTEIN SERINE THREONINE KINASE; TISSUE EXTRACT; XENOPUS PROTEIN;

ANIMAL; ARTICLE; CELL FREE SYSTEM; CHEMISTRY; CHROMATIN; DNA DAMAGE; DRUG EFFECT; FEMALE; GENETICS; ISOLATION AND PURIFICATION; MALE; METABOLISM; OOCYTE; RADIATION EXPOSURE; SPERMATOZOON; ULTRAVIOLET RADIATION; XENOPUS LAEVIS;

ANIMALS; CELL CYCLE PROTEINS; CELL-FREE SYSTEM; CHROMATIN; DNA; DNA DAMAGE; FEMALE; MALE; MITOMYCIN; OVUM; PROTEIN KINASES; PROTEIN-SERINE-THREONINE KINASES; SPERMATOZOA; TISSUE EXTRACTS; ULTRAVIOLET RAYS; XENOPUS LAEVIS; XENOPUS PROTEINS;

EID: 84869219091     PISSN: 1940087X     EISSN: None     Source Type: Journal    
DOI: 10.3791/4449     Document Type: Article

References (14)

1. Lohka, M.J. & Masui, Y. Formation in vitro of sperm pronuclei and mitotic chromosomes induced by amphibian ooplasmic components. Science. 220 (4598), 719-721 (1983).
2. Cimprich, K.A. & Cortez, D. ATR: an essential regulator of genome integrity. Nat. Rev. Mol. Cell Biol. 9 (8), 616-627 (2008).
3. Lupardus, P.J., Van, C., & Cimprich, K.A. Analyzing the ATR-mediated checkpoint using Xenopus egg extracts. Methods. 41 (2), 222-231 (2007).
4. Lupardus, P.J., Byun, T., Yee, M.C., Hekmat-Nejad, M., & Cimprich, K.A. A requirement for replication in activation of the ATR-dependent DNA damage checkpoint. Genes Dev. 16 (18), 2327-2332 (2002).
5. Stokes, M.P., Van Hatten, R., Lindsay, H.D., & Michael, W.M. DNA replication is required for the checkpoint response to damaged DNA in Xenopus egg extracts. J. Cell Biol. 158 (5), 863-872 (2002).
6. Kato, K. & Strauss, B. Accumulation of an intermediate in DNA synthesis by HEp.2 cells treated with methyl methanesulfonate. Proc. Natl. Acad. Sci. U.S.A. 71 (5), 1969-1973 (1974).
7. Paulovich, A.G. & Hartwell, L.H. A checkpoint regulates the rate of progression through S phase in S. cerevisiae in response to DNA damage. Cell. 82 (5), 841-847 (1995).
8. Guo, Z., Kumagai, A., Wang, S.X., & Dunphy, W. G. Requirement for Atr in phosphorylation of Chk1 and cell cycle regulation in response to DNA replication blocks and UV-damaged DNA in Xenopus egg extracts. Genes Dev. 14 (21), 2745-2756 (2000).
9. Jazayeri, A., Balestrini, A., Garner, E., Haber, J.E., & Costanzo, V. Mre11-Rad50-Nbs1-dependent processing of DNA breaks generates oligonucleotides that stimulate ATM activity. EMBO. J. 27 (14), 1953-1962 (2008).
10. Kumagai, A. & Dunphy, W.G. Claspin, a novel protein required for the activation of Chk1 during a DNA replication checkpoint response in Xenopus egg extracts. Mol. Cell. 6 (4), 839-849 (2000).
11. Yan, S., Lindsay, H.D., & Michael, W.M. Direct requirement for Xmus101 in ATR-mediated phosphorylation of Claspin bound Chk1 during checkpoint signaling. J. Cell Biol. 173 (2), 181-186 (2006).
12. Shiotani, B. & Zou, L. Single-stranded DNA orchestrates an ATM-to-ATR switch at DNA breaks. Mol. Cell. 33 (5), 547-558 (2009).
13. Tutter, A.V. & Walter, J.C. Chromosomal DNA replication in a soluble cell-free system derived from Xenopus eggs. Methods Mol. Biol. 322, 121-137 (2006).
14. Cross, M.K. & Powers, M. Preparation and Fractionation of Xenopus laevis Egg Extracts. J. Vis. Exp. (18), e891, DOI: 10.3791/891 (2008).