Prokaryotic BirA ligase biotinylates K4, K9, K18 and K23 in histone H3.
BirA ligase is a prokaryotic ortholog of holocarboxylase synthetase (HCS) that can biotinylate proteins. This study tested the hypothesis that BirA ligase catalyzes the biotinylation of eukaryotic histones. If so, this would mean that recombinant BirA ligase is a useful surrogate for HCS in studies of histone biotinylation. The biological activity of recombinant BirA ligase was confirmed by enzymatic biotinylation of p67. In particular, it was found that BirA ligase biotinylated both calf thymus histone H1 and human bulk histone extracts. Incubation of recombinant BirA ligase with H3-based synthetic peptides showed that lysines 4, 9, 18, and 23 in histone H3 are the targets for the biotinylation by BirA ligase. Modification of the peptides (e.g., serine phosphorylation) affected the subsequent biotinylation by BirA ligase, suggesting crosstalk between modifications. In conclusion, this study suggests that prokaryotic BirA ligase is a promiscuous enzyme and biotinylates eukaryotic histones. Moreover the biotinylation of histones by BirA ligase is consistent with the proposed role of human HCS in chromatin.
Kobza
Keyna
K
Departments of Nutrition and Health Sciences, University of Nebraska at Lincoln, Lincoln, NE, USA.
Sarath
Gautam
G
Zempleni
Janos
J
D000595:Amino Acid Sequence; D019921:Biotinylation; D019731:Carbon-Nitrogen Ligases / Q000378:metabolism / Q000502:physiology*; D002843:Chromatin / Q000378:metabolism; D029968:Escherichia coli Proteins / Q000378:metabolism / Q000502:physiology*; D006657:Histones / Q000737:chemistry / Q000378:metabolism*; D006801:Humans; D019169:Jurkat Cells; D008025:Ligases / Q000378:metabolism / Q000502:physiology; D008239:Lysine / Q000378:metabolism*; D008969:Molecular Sequence Data; D012097:Repressor Proteins / Q000378:metabolism / Q000502:physiology*; D014157:Transcription Factors / Q000378:metabolism / Q000502:physiology*
D002843:Chromatin; D029968:Escherichia coli Proteins; D006657:Histones; D012097:Repressor Proteins; D014157:Transcription Factors; D008025:Ligases; D019731:Carbon-Nitrogen Ligases; C023312:holocarboxylase synthetases; C058125:birA protein, E coli; D008239:Lysine