Replacement of 2′-Deoxycytidine by 2′-Deoxycytidine Analogues in the E. coli Genome

Angad P. Mehta, Han Li, Sean A. Reed, Lubica Supekova, Tsotne Javahishvili, Peter G. Schultz

Research output: Contribution to journalArticlepeer-review


Several modified bases have been observed in the genomic DNA of bacteriophages, prokaryotes, and eukaryotes that play a role in restriction systems and/or epigenetic regulation. In our efforts to understand the consequences of replacing a large fraction of a canonical nucleoside with a modified nucleoside, we previously replaced around 75% of thymidine (T) with 5′-hydroxymethyl-2′-deoxyuridine (5hmU) in the Escherichia coli genome. In this study, we engineered the pyrimidine nucleotide biosynthetic pathway using T4 bacteriophage genes to achieve approximately 63% replacement of 2′-deoxycytidine (dC) with 5-hydroxymethyl-2′-deoxycytidine (5hmC) in the E. coli genome and approximately 71% replacement in plasmids. We further engineered the glucose metabolic pathway to transform the 5hmC into glucosyl-5-hydroxymethyl-2′-deoxycytidine (5-gmC) and achieved 20% 5-gmC in the genome and 45% 5-gmC in plasmid DNA.

Original languageEnglish (US)
Pages (from-to)14230-14233
Number of pages4
JournalJournal of the American Chemical Society
Issue number43
StatePublished - Nov 2 2016
Externally publishedYes

ASJC Scopus subject areas

  • Catalysis
  • General Chemistry
  • Biochemistry
  • Colloid and Surface Chemistry


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