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Ligase-Mediated Threose Nucleic Acid Synthesis on DNA Templates.
ACS Synth Biol 2019; 8(2):282-286AS

Abstract

Ligases are a class of enzymes that catalyze the formation of phosphodiester bonds between an oligonucleotide donor with a 5' terminal phosphate and an oligonucleotide acceptor with a 3' terminal hydroxyl group. Here, we wished to explore the substrate specificity of naturally occurring DNA and RNA ligases to determine whether the molecular recognition of these enzymes is sufficiently general to synthesize alternative genetic polymers with backbone structures that are distinct from those found in nature. We chose threose nucleic acid (TNA) as a model system, as TNA is known to be biologically stable and capable of undergoing Darwinian evolution. Enzyme screening and reaction optimization identified several ligases that can recognize TNA as either the donor or acceptor strand with DNA. Less discrimination occurs on the acceptor strand indicating that the determinants of substrate specificity depend primarily on the composition of the donor strand. Remarkably, T3 and T7 ligases were able to join TNA homopolymers together, which is surprising given that the TNA backbone is one atom shorter than that of DNA. In this reaction, the base composition of the ligation junction strongly favors the formation of A-T and A-G linkages. We suggest that these results will enable the assembly of TNA oligonucleotides of lengths beyond what is currently possible by solid-phase synthesis and provide a starting point for further optimization by directed evolution.

Authors

No affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, U.S. Gov't, Non-P.H.S.

Language

eng

PubMed ID

30629885

Citation

McCloskey, Cailen M., et al. "Ligase-Mediated Threose Nucleic Acid Synthesis On DNA Templates." ACS Synthetic Biology, vol. 8, no. 2, 2019, pp. 282-286.
McCloskey CM, Liao JY, Bala S, et al. Ligase-Mediated Threose Nucleic Acid Synthesis on DNA Templates. ACS Synth Biol. 2019;8(2):282-286.
McCloskey, C. M., Liao, J. Y., Bala, S., & Chaput, J. C. (2019). Ligase-Mediated Threose Nucleic Acid Synthesis on DNA Templates. ACS Synthetic Biology, 8(2), pp. 282-286. doi:10.1021/acssynbio.8b00511.
McCloskey CM, et al. Ligase-Mediated Threose Nucleic Acid Synthesis On DNA Templates. ACS Synth Biol. 2019 02 15;8(2):282-286. PubMed PMID: 30629885.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Ligase-Mediated Threose Nucleic Acid Synthesis on DNA Templates. AU - McCloskey,Cailen M, AU - Liao,Jen-Yu, AU - Bala,Saikat, AU - Chaput,John C, Y1 - 2019/01/14/ PY - 2019/1/11/pubmed PY - 2019/1/11/medline PY - 2019/1/11/entrez KW - enzymatic ligation KW - ligase KW - threose nucleic acid KW - xeno nucleic acid SP - 282 EP - 286 JF - ACS synthetic biology JO - ACS Synth Biol VL - 8 IS - 2 N2 - Ligases are a class of enzymes that catalyze the formation of phosphodiester bonds between an oligonucleotide donor with a 5' terminal phosphate and an oligonucleotide acceptor with a 3' terminal hydroxyl group. Here, we wished to explore the substrate specificity of naturally occurring DNA and RNA ligases to determine whether the molecular recognition of these enzymes is sufficiently general to synthesize alternative genetic polymers with backbone structures that are distinct from those found in nature. We chose threose nucleic acid (TNA) as a model system, as TNA is known to be biologically stable and capable of undergoing Darwinian evolution. Enzyme screening and reaction optimization identified several ligases that can recognize TNA as either the donor or acceptor strand with DNA. Less discrimination occurs on the acceptor strand indicating that the determinants of substrate specificity depend primarily on the composition of the donor strand. Remarkably, T3 and T7 ligases were able to join TNA homopolymers together, which is surprising given that the TNA backbone is one atom shorter than that of DNA. In this reaction, the base composition of the ligation junction strongly favors the formation of A-T and A-G linkages. We suggest that these results will enable the assembly of TNA oligonucleotides of lengths beyond what is currently possible by solid-phase synthesis and provide a starting point for further optimization by directed evolution. SN - 2161-5063 UR - https://www.unboundmedicine.com/medline/citation/30629885/Ligase_Mediated_Threose_Nucleic_Acid_Synthesis_on_DNA_Templates_ L2 - https://dx.doi.org/10.1021/acssynbio.8b00511 DB - PRIME DP - Unbound Medicine ER -