Post-synthetic acetylation of HMGB1 protein modulates its interactions with supercoiled DNA
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High mobility group box (HMGB) proteins 1 and 2 are abundant non-histone nuclear proteins that regulate chromatin structure because of their structure-specific binding to DNA. Here, we have investigated how the post-synthetic acetylation of HMGB1 affects its interaction with negatively supercoiled DNA by employing monoacetylated at Lys2 protein, isolated from butyrate-treated cells. Our data reveal that this modification enhances three reaction parameters: binding affinity, supercoiling activity and capacity to protect the supercoiled DNA from relaxation by topoisomerase I. We show that monoacetylation at Lys2 mimics the effect of acidic tail removal but to a lesser extent thus demonstrating that in vivo acetylated HMGB1 is capable of modulating its interaction with negatively supercoiled DNA.
KeywordsAcetylated HMGB1 HMGB1 protein Supercoiled DNA Tailless HMGB1
Electrophoretic mobility shift assay
High mobility group box protein 1
Truncated HMGB1 protein, lacking the acidic tail
Recombinant HMGB1 protein
Monoacetylated HMGB1 protein
- S, L
Supercoiled and linearized forms of pEGFP-N, respectively
This work was partially supported by Grant TKB 1608 from the National Science Fund, Ministry of Education and Science of Republic of Bulgaria.
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