title: Direct conjugation of peptides and 5-hydroxymethylcytosine in DNA.
abstract: Recent discovery of functional 5-hydroxymethylcytosine in vertebrate genomes prompted for elaboration of methods to localize this modification at the nucleotide resolution level. Among several covalent modification-based approaches, atypical activity of cytosine-5 DNA methyltransferases to couple small molecules to 5-hydroxymethylcytosine stands out for acceptance of broad range of ligands. We went further to explore the possibility for methyltransferase-maintained coupling of compounds possessing autonomous functions. Functionalization of DNA was achieved by direct conjugation of chemically synthesized peptides of regular structure. Sequence, residue, and position-specific coupling of DNA containing 5-hydroxymethylcytosine and different peptides has been demonstrated, with the nature of the resulting conjugates confirmed by protease treatment and mass spectrometry. Coupling products were compatible with affinity-driven separation from the unmodified DNA. This approach highlights an emerging avenue toward the enzymatic, sequence-specific DNA functionalization, enabling a single step merge of the DNA and peptide moieties into a bifunctional entity.
authors.affiliation: †Thermo Fisher Scientific Baltics, Graičiu̅no 8, Vilnius LT-02241, Lithuania.|‡Department of Biochemistry and Molecular Biology, Faculty of Natural Sciences, Vilnius University, MK Čiurlionio 21/27, LT-03101 Vilnius, Lithuania.
authors.forename: Saulius
authors.initials: S
authors.lastname: Serva
authors.affiliation: †Thermo Fisher Scientific Baltics, Graičiu̅no 8, Vilnius LT-02241, Lithuania.
authors.forename: Arūnas
authors.initials: A
authors.lastname: Lagunavičius
mesh_terms: D044503:5-Methylcytosine / Q000031:analogs & derivatives; D000595:Amino Acid Sequence; D003596:Cytosine / Q000031:analogs & derivatives* / Q000138:chemical synthesis / Q000737:chemistry / Q000378:metabolism; D004247:DNA / Q000737:chemistry* / Q000378:metabolism; D015257:DNA-Cytosine Methylases / Q000378:metabolism; D008958:Models, Molecular; D010455:Peptides / Q000138:chemical synthesis / Q000737:chemistry* / Q000378:metabolism; D013149:Spiroplasma / Q000201:enzymology
chemical_list: D010455:Peptides; C011865:5-hydroxymethylcytosine; D044503:5-Methylcytosine; D003596:Cytosine; D004247:DNA; C037664:DNA modification methylase HhaI; C081573:DNA modification methylase SssI; D015257:DNA-Cytosine Methylases