Acetylation regulates DNA repair mechanisms in human cells

The p300-mediated acetylation of enzymes involved in DNA repair and replication has been previously shown to stimulate or inhibit their activities in reconstituted systems. To explore the role of acetylation on DNA repair in cells we constructed plasmid substrates carrying inactivating damages in th...

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Bibliographic Details
Published in:Cell cycle (Georgetown, Tex.) Tex.), 2016-06, Vol.15 (11), p.1506-1517
Main Authors: Piekna-Przybylska, Dorota, Bambara, Robert A, Balakrishnan, Lata
Format: Article
Language:English
Subjects:
Acetylation
Amino Acid Sequence
Base Pair Mismatch
Base Sequence
DNA - genetics
DNA - metabolism
DNA Mismatch Repair
DNA Repair
DNA Replication
Enzyme Inhibitors - pharmacology
Genes, Reporter
Green Fluorescent Proteins - genetics
Green Fluorescent Proteins - metabolism
Guanine - analogs & derivatives
Guanine - metabolism
HCT116 Cells
HeLa Cells
Humans
p300-CBP Transcription Factors - genetics
p300-CBP Transcription Factors - metabolism
Plasmids - chemistry
Plasmids - metabolism
Protein Processing, Post-Translational
Sequence Alignment
Terpenes - pharmacology
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Summary:The p300-mediated acetylation of enzymes involved in DNA repair and replication has been previously shown to stimulate or inhibit their activities in reconstituted systems. To explore the role of acetylation on DNA repair in cells we constructed plasmid substrates carrying inactivating damages in the EGFP reporter gene, which should be repaired in cells through DNA mismatch repair (MMR) or base excision repair (BER) mechanisms. We analyzed efficiency of repair within these plasmid substrates in cells exposed to deacetylase and acetyltransferase inhibitors, and also in cells deficient in p300 acetyltransferase. Our results indicate that protein acetylation improves DNA mismatch repair in MMR-proficient HeLa cells and also in MMR-deficient HCT116 cells. Moreover, results suggest that stimulated repair of mismatches in MMR-deficient HCT116 cells is done though a strand-displacement synthesis mechanism described previously for Okazaki fragments maturation and also for the EXOI-independent pathway of MMR. Loss of p300 reduced repair of mismatches in MMR-deficient cells, but did not have evident effects on BER mechanisms, including the long patch BER pathway. Hypoacetylation of the cells in the presence of acetyltransferase inhibitor, garcinol generally reduced efficiency of BER of 8-oxoG damage, indicating that some steps in the pathway are stimulated by acetylation.
ISSN:1538-4101
1551-4005
DOI:10.1080/15384101.2016.1176815