Abstract
Induction of DNA double strand breaks leads to phosphorylation and focus-formation of H2AX. However, foci of phosphorylated H2AX (γH2AX) appear during DNA replication also in the absence of exogenously applied injury. We measured the amount and the number of foci of γH2AX in different phases of the cell cycle by flow cytometry, sorting and microscopy in 4 malignant B-lymphocyte cell lines. There were no detectable γH2AX and no γH2AX-foci in G1 cells in exponentially growing cells and cells treated with PARP inhibitor (PARPi) for 24 h to create damage and reduce DNA repair. The amount of γH2AX increased immediately upon S phase entry, and about 10 and 30 γH2AX foci were found in mid-S phase control and PARPi-treated cells, respectively. The γH2AX-labeled damage caused by DNA replication was not fully repaired before entry into G2. Intriguingly, G2 cells populated a continuous distribution of γH2AX levels, from cells with a high content of γH2AX and the same number of foci as S phase cells (termed "G2H" compartment), to cells that there were almost negative and had about 2 foci (termed "G2L" compartment). EdU-labeling of S phase cells revealed that G2H was directly populated from S phase, while G2L was populated from G2H, but in ...Continue Reading
References
Dec 1, 1975·Proceedings of the National Academy of Sciences of the United States of America·A B Pardee, L J James
Aug 15, 1992·Proceedings of the National Academy of Sciences of the United States of America·S J KuerbitzM B Kastan
Jun 19, 2001·Biochimica Et Biophysica Acta·V A Smits, R H Medema
Nov 14, 2002·Cell Cycle·Mikhail V Blagosklonny, Arthur B Pardee
Apr 5, 2003·Cancer Cell·Claus Storgaard SørensenJiri Lukas
May 20, 2003·Radiation Research·Susan H MacPhailPeggy L Olive
Jun 11, 2004·Cell Cycle·Claus Storgaard SørensenJiri Bartek
Jul 29, 2004·DNA Repair·Jiri LukasJiri Bartek
Sep 28, 2004·Radiation Research·Kirsti Solberg LandsverkTrond Stokke
Jul 21, 2005·Molecular Biology of the Cell·Kirk J McManus, Michael J Hendzel
Aug 16, 2005·Cell Proliferation·Xuan HuangZbigniew Darzynkiewicz
Sep 13, 2005·Biochemical and Biophysical Research Communications·Yosuke IchijimaHirobumi Teraoka
Jan 11, 2007·British Journal of Cancer·M Damelin, T H Bestor
Oct 19, 2007·Mathematical Biosciences·Sara LarssonMaria Johansson
Feb 9, 2010·Cell Cycle·Markus LöbrichPenny A Jeggo
Jul 5, 2011·Radiotherapy and Oncology : Journal of the European Society for Therapeutic Radiology and Oncology·Kinga Tkacz-StachowskaRandi G Syljuåsen
Nov 3, 2012·Cancer Research·Junko MuraiYves Pommier
Jun 15, 2014·Strahlentherapie und Onkologie : Organ der Deutschen Röntgengesellschaft ... [et al]·Friedo ZölzerChristian Streffer
Sep 10, 2014·Journal of Molecular Biology·Jonathan Baxter
Citations
Jun 12, 2018·PloS One·Masaki ShigetaTakahiko Yokoyama
Sep 5, 2019·British Journal of Cancer·Nicholas R JetteSusan P Lees-Miller
Mar 23, 2017·Stem Cell Research & Therapy·Pavel SimaraIrena Koutna
Jan 17, 2020·Cytometry. Part a : the Journal of the International Society for Analytical Cytology·Idun D ReinTrond Stokke
Mar 20, 2021·Scientific Reports·Morten P OksvoldThea Kristin Våtsveen