Ellagic Acid-Changed Epigenome of Ribosomal Genes and Condensed RPA194-Positive Regions of Nucleoli in Tumour Cells.
We studied the effect of ellagic acid (EA) on the morphology of nucleoli and on the pattern of major proteins of the nucleolus. After EA treatment of HeLa cells, we observed condensation of nucleoli as documented by the pattern of argyrophilic nucleolar organizer regions (AgNORs). EA also induced condensation of RPA194-positive nucleolar regions, but no morphological changes were observed in nucleolar compartments positive for UBF1/2 proteins or fibrillarin. Studied morphological changes induced by EA were compared with the morphology of control, non-treated cells and with pronounced condensation of all nucleolar domains caused by actinomycin D (ACT-D) treatment. Similarly as ACT-D, but in a lesser extent, EA induced an increased number of 53BP1-positive DNA lesions. However, the main marker of DNA lesions, γH2AX, was not accumulated in body-like nuclear structures. An increased level of γH2AX was found by immunofluorescence and Western blots only after EA treatment. Intriguingly, the levels of fibrillarin, UBF1/2 and γH2AX were increased at the promoters of ribosomal genes, while 53BP1 and CARM1 levels were decreased by EA treatment at these genomic regions. In the entire genome, EA reduced H3R17 dimethylation. Taken together, ellagic acid is capable of significantly changing the nucleolar morphology and protein levels inside the nucleolus.
Institute of Biophysics, Academy of Sciences of the Czech Republic, v. v. i., Brno, Czech Republic.
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Legartová
Epigenetic MedChem Lab, Università di Salerno Dipartimento di Farmacia, Fisciano, Salerno, Italy.
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Sbardella
Institute of Biophysics, Academy of Sciences of the Czech Republic, v. v. i., Brno, Czech Republic.
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Kozubek
Institute of Biophysics, Academy of Sciences of the Czech Republic, v. v. i., Brno, Czech Republic.
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Bártová
D053468:CARD Signaling Adaptor Proteins / Q000032:analysis / Q000037:antagonists & inhibitors*; D002455:Cell Division / Q000187:drug effects; D002466:Cell Nucleolus / Q000737:chemistry / Q000187:drug effects* / Q000648:ultrastructure; D002868:Chromosomal Proteins, Non-Histone / Q000032:analysis; D004249:DNA Damage; D004275:DNA, Ribosomal / Q000187:drug effects* / Q000235:genetics; D003609:Dactinomycin / Q000494:pharmacology; D004610:Ellagic Acid / Q000494:pharmacology*; D044127:Epigenesis, Genetic / Q000187:drug effects*; D016195:G2 Phase / Q000187:drug effects; D006162:Guanylate Cyclase / Q000032:analysis / Q000037:antagonists & inhibitors*; D006367:HeLa Cells / Q000737:chemistry / Q000187:drug effects; D006657:Histones / Q000032:analysis / Q000378:metabolism; D006801:Humans; D047908:Intracellular Signaling Peptides and Proteins / Q000032:analysis; D008745:Methylation; D009363:Neoplasm Proteins / Q000032:analysis; D009697:Nucleolus Organizer Region / Q000737:chemistry / Q000187:drug effects / Q000648:ultrastructure; D035463:Pol1 Transcription Initiation Complex Proteins / Q000032:analysis; D011401:Promoter Regions, Genetic; D011499:Protein Processing, Post-Translational / Q000187:drug effects*; D012318:RNA Polymerase I / Q000032:analysis; D000071857:Tumor Suppressor p53-Binding Protein 1
D053468:CARD Signaling Adaptor Proteins; D002868:Chromosomal Proteins, Non-Histone; D004275:DNA, Ribosomal; C468783:H2AX protein, human; D006657:Histones; D047908:Intracellular Signaling Peptides and Proteins; D009363:Neoplasm Proteins; D035463:Pol1 Transcription Initiation Complex Proteins; C498301:TP53BP1 protein, human; D000071857:Tumor Suppressor p53-Binding Protein 1; C047458:fibrillarin; C063844:transcription factor UBF; D004610:Ellagic Acid; D003609:Dactinomycin; C000608127:RPA194 protein, human; D012318:RNA Polymerase I; C426242:CARD11 protein, human; D006162:Guanylate Cyclase