caa a22 4500 463160010 CHVBK 20180406164744.0 cr unu---uuuuu 170326e20070901xx s 000 0 eng 10.1007/s10535-007-0094-9 doi (NATIONALLICENCE)springer-10.1007/s10535-007-0094-9 Induction of foci of phosphorylated H2AX histones and premature chromosome condensation after DNA damage in Vicia faba root meristem [Elektronische Daten] [D. Rybaczek, J. Maszewski] Immunocytochemical analysis using antibody raised against human H2AX histones phosphorylated at serine 139 (γ-H2AX) demonstrates that root meristem cells of Vicia faba exposed to UV-radiation or incubated with hydroxyurea (HU) reveal discrete foci at the border of the nucleolus and perinucleolar chromatin or scattered over the whole area of cell nucleus. Western blots detected only one protein band at the position expected for the phosphorylated form of H2AX. The dose-effect relationship was demonstrated following treatment with 2.5 and 10 mM HU. Proteins extracted from root meristems incubated for 2 h either with HU and caffeine or with HU and sodium metavanadate showed unchanged amounts of bound γ-H2AX antibodies, as compared to root meristems treated with 2.5 mM HU. Higher quantities of phosphorylated H2AX histones were detected in proteins extracted from roots treated with HU and 2-aminopurine. All treatments were effective in producing evident aberrations of premature mitosis: broken and lagging chromatids, acentric fragments, chromosomal bridges and micronuclei. Our results show that phosphorylation of H2AX at the carboxy-terminal Ser-Gln-Glu sequence is among the earliest responses to double-strand breaks and, presumably, one of the key ATM/ATR-dependent signals indispensable for the repair of spontaneous and induced DNA damage in plant cells. Institute of Experimental Botany, ASCR, 2007 field bean nationallicence hydroxyurea nationallicence premature mitosis nationallicence UV-radiation nationallicence AP : 2-aminopurine nationallicence ATM : ataxia telangiectasia mutated nationallicence ATR : ATM and Rad3-related nationallicence CF : caffeine nationallicence dNTPs : deoxyribonucleoside triphosphates nationallicence DSB : double-strand breaks nationallicence HU : hydroxyurea nationallicence IR : ionizing radiation nationallicence MMS : methyl methane sulfonate nationallicence MPF : M-phase promoting factor nationallicence PCC : premature chromosome condensation nationallicence PCNA : proliferating cell nuclear antigen nationallicence PI3K : phosphoinositide 3-kinase-like kinase nationallicence RNR : ribonucleotide reductase nationallicence ssDNA : single-stranded DNA nationallicence Van : sodium metavanadate nationallicence Rybaczek D. Department of Cytophysiology, University of Łódź, ul. Pilarskiego 14, PL-90231, Łódź, Poland aut Maszewski J. Department of Cytophysiology, University of Łódź, ul. Pilarskiego 14, PL-90231, Łódź, Poland aut Biologia Plantarum Kluwer Academic Publishers-Consultants Bureau 51/3(2007-09-01), 443-450 0006-3134 51:3<443 2007 51 10535 https://doi.org/10.1007/s10535-007-0094-9 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1007/s10535-007-0094-9 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Rybaczek D. Department of Cytophysiology, University of Łódź, ul. Pilarskiego 14, PL-90231, Łódź, Poland aut NATIONALLICENCE 700 1- Maszewski J. Department of Cytophysiology, University of Łódź, ul. Pilarskiego 14, PL-90231, Łódź, Poland aut NATIONALLICENCE 773 0- Biologia Plantarum Kluwer Academic Publishers-Consultants Bureau 51/3(2007-09-01), 443-450 0006-3134 51:3<443 2007 51 10535 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer