SILAC-based quantification of changes in chromatin association in cells lacking Rrm3.
<p>(A) Equal numbers of light-labeled (wildtype) and heavy-labeled (<i>rrm3Δ</i>) cells were mixed, nuclei isolated, and the chromatin fraction extracted and analyzed by tandem mass spectrometry using a hybrid linear ion trap-Orbitrap instrument. (B) Subcellular fractionation was verified by following the distribution of proteins in cytoplasmic (Cyto), nucleoplasm (NP), and chromatin (Ch) fractions during the enrichment procedure by Western blotting. Successful chromatin fractionation is indicated by the enrichment of histone H3 and Rfa and the absence of Adh1. Chromatin fractions were analyzed from three biological replicates in (C) the absence of hydroxyurea and (D) in the presence of hydroxyurea. (E) Merger of peptide quantification in the absence and presence of hydroxyurea.</p
Biophysics, Biochemistry, Cell Biology, Genetics, Molecular Biology, Developmental Biology, Cancer, Science Policy, Environmental Sciences not elsewhere classified, Orc 5-Binding Domain, Rrm 3 function, replication stressor hydroxyurea, replication checkpoint mediator Mrc 1, Rrm 3 associates, increase nucleotide levels, replication stress cells, binding Orc 5, DNA repair pathways, Restricting DNA Replication, novel Rrm 3 function, DNA damage tolerance, replication stress, Novel Rrm 3 Function, DNA synthesis maps, Orc 5-binding domain, Facilitating Fork Progression, controls DNA synthesis, Rrm 3 Function, DNA synthesis, replication fork progression, recombination factor Rdh 54, DNA lesions
DOI identifier: 10.1371/journal.pgen.1006451.g001
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