Replacement of 2'-Deoxycytidine by 2'-Deoxycytidine Analogues in the E. Coli Genome

Overview

Journal J Am Chem Soc

Publisher American Chemical Society

Specialty Chemistry

Date 2016 Nov 3

PMID 27762133

Citations 9

Authors

Angad P Mehta

Han Li

Sean A Reed

Lubica Supekova

Tsotne Javahishvili

Peter G Schultz

Affiliations

Soon will be listed here.

Abstract

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.

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