The overwhelmingly homochiral nature of life has left a puzzle as to whether mirror-image biological systems based on a chirally inverted version of molecular machinery could also have existed. Here we report that two key steps in the central dogma of molecular biology, the template-directed polymerization of DNA and transcription into RNA, can be catalysed by a chemically synthesized D-amino acid polymerase on an L-DNA template. We also show that two chirally mirrored versions of the 174-residue African swine fever virus polymerase X could operate in a racemic mixture without significant enantiomeric cross-inhibition to the activity of each other. Furthermore, we demonstrate that a functionally active L-DNAzyme could be enzymatically produced using the D-amino acid polymerase. The establishment of such molecular systems with an opposite handedness highlights the potential to exploit enzymatically produced mirror-image biomolecules as research and therapeutic tools.
Subscribe to Journal
Get full journal access for 1 year
only $14.08 per issue
All prices are NET prices.
VAT will be added later in the checkout.
Rent or Buy article
Get time limited or full article access on ReadCube.
All prices are NET prices.
Imai, K. et al. Analytical chemistry and biochemistry of D-amino acids. Biomed. Chromatogr. 10, 303–312 (1996).
Northcote, D. Chemistry of the plant cell wall. Ann. Rev. Plant Physiol. 23, 113–132 (1972).
Islas, J. R. et al. Mirror-symmetry breaking in the Soai reaction: a kinetic understanding. Proc. Natl Acad. Sci. USA 102, 13743–13748 (2005).
Glavin, D. P. & Dworkin, J. P. Enrichment of the amino acid L-isovaline by aqueous alteration on CI and CM meteorite parent bodies. Proc. Natl Acad. Sci. USA 106, 5487–5492 (2009).
Kondepudi, D. K., Kaufman, R. J. & Singh, N. Chiral symmetry breaking in sodium chlorate crystallization. Science 250, 975–976 (1990).
Cordova, A., Engqvist, M., Ibrahem, I., Casas, J. & Sunden, H. Plausible origins of homochirality in the amino acid catalyzed neogenesis of carbohydrates. Chem. Commun. 2047–2049 (2005).
Sczepanski, J. T. & Joyce, G. F. A cross-chiral RNA polymerase ribozyme. Nature 515, 440–442 (2014).
Bohannon, J. Mirror-image cells could transform science—or kill us all. Wired (2010); http://www.wired.com/2010/11/ff_mirrorlife/
Milton, R., Milton, S. & Kent, S. Total chemical synthesis of a D-enzyme: the enantiomers of HIV-1 protease show reciprocal chiral substrate specificity. Science 256, 1445–1448 (1992).
Wyszko, E. et al. Spiegelzymes: sequence specific hydrolysis of L-RNA with mirror image hammerhead ribozymes and DNAzymes. PloS ONE 8, e54741 (2013).
Urata, H., Ogura, E., Shinohara, K., Ueda, Y. & Akagi, M. Synthesis and properties of mirror-image DNA. Nucleic Acids Res. 20, 3325–3332 (1992).
Wyszko, E. et al. Spiegelzymes mirror-image hammerhead ribozymes and mirror-image DNAzymes, an alternative to siRNAs and microRNAs to cleave mRNAs. PloS ONE 9, e86673 (2014).
Kent, S. B. H. Total chemical synthesis of proteins. Chem. Soc. Rev. 38, 338–351 (2009).
Zheng, J.-S., Tang, S., Qi, Y.-K., Wang, Z.-P. & Liu, L. Chemical synthesis of proteins using peptide hydrazides as thioester surrogates. Nature Protocols 8, 2483–2495 (2013).
Weinstock, M. T., Jacobsen, M. T. & Kay, M. S. Synthesis and folding of a mirror-image enzyme reveals ambidextrous chaperone activity. Proc. Natl Acad. Sci. USA 111, 11679–11684 (2014).
Oliveros, M. et al. Characterization of an African swine fever virus 20-kDa DNA polymerase involved in DNA repair. J. Biol. Chem. 272, 30899–30910 (1997).
Vinogradov, A. A., Evans, E. D. & Pentelute, B. L. Total synthesis and biochemical characterization of mirror image barnase. Chem. Sci. 6, 2997–3002 (2015).
Fang, G.-M. et al. Protein chemical synthesis by ligation of peptide hydrazides. Angew. Chem. Int. Ed. 50, 7645–7649 (2011).
Tang, S. et al. An efficient one-pot four-segment condensation method for protein chemical synthesis. Angew. Chem. Int. Ed. 54, 5713–5717 (2015).
Coin, I. The depsipeptide method for solid-phase synthesis of difficult peptides. J. Peptide Sci. 16, 223–230 (2010).
Liu, F., Luo, E. Y., Flora, D. B. & Mezo, A. R. A synthetic route to human insulin using isoacyl peptides. Angew. Chem. Int. Ed. 53, 3983–3987 (2014).
Williams, K. P. et al. Bioactive and nuclease-resistant L-DNA ligand of vasopressin. Proc. Natl Acad. Sci. USA 94, 11285–11290 (1997).
Showalter, A. K. & Tsai, M. D. A DNA polymerase with specificity for five base pairs. J. Am. Chem. Soc. 123, 1776–1777 (2001).
Yamaguchi, T., Iwanami, N., Shudo, K. & Saneyoshi, M. Chiral discrimination of enantiomeric 2ʹ-deoxythymidine 5ʹ-triphosphate by HIV-1 reverse transcriptase and eukaryotic DNA polymerases. Biochem. Biophys. Res. Commun. 200, 1023–1027 (1994).
Sosunov, V. V. et al. Stereochemical control of DNA biosynthesis. Nucleic Acids Res. 28, 1170–1175 (2000).
Semizarov, D. G. et al. Stereoisomers of deoxynucleoside 5ʹ-triphosphates as substrates for template-dependent and -independent DNA polymerases. J. Biol. Chem. 272, 9556–9560 (1997).
Focher, F. et al. Stereospecificity of human DNA polymerases α, β, γ, δ and ε, HIV-reverse transcriptase, HSV-1 DNA polymerase, calf thymus terminal transferase and Escherichia coli DNA polymerase I in recognizing D- and L-thymidine 5ʹ-triphosphate as substrate. Nucleic Acids Res. 23, 2840–2847 (1995).
Siegel, J. S. Left-handed comments. Science 258, 1290 (1992).
Gu, H., Furukawa, K., Weinberg, Z., Berenson, D. F. & Breaker, R. R. Small, highly active DNAs that hydrolyze DNA. J. Am. Chem. Soc. 135, 9121–9129 (2013).
Yatime, L. et al. Structural basis for the targeting of complement anaphylatoxin C5a using a mixed L-RNA/L-DNA aptamer. Nature Commun. 6, 6481 (2015).
Boudsocq, F., Iwai, S., Hanaoka, F. & Woodgate, R. Sulfolobus solfataricus P2 DNA polymerase IV (Dpo4): an archaeal DinB-like DNA polymerase with lesion-bypass properties akin to eukaryotic polη. Nucleic Acids Res. 29, 4607–4616 (2001).
Nick McElhinny, S. A. & Ramsden, D. A. Polymerase mu is a DNA-directed DNA/RNA polymerase. Mol. Cell. Biol. 23, 2309–2315 (2003).
Pinheiro, V. B. et al. Synthetic genetic polymers capable of heredity and evolution. Science 336, 341–344 (2012).
Jewett, M. C., Fritz, B. R., Timmerman, L. E. & Church, G. M. In vitro integration of ribosomal RNA synthesis, ribosome assembly, and translation. Mol. Syst. Biol. 9, 678 (2013).
Stelzl, U., Connell, S., Nierhaus, K. H. & Wittmann-Liebold, B. Ribosomal proteins: role in ribosomal functions. eLS http://dx.doi.org/10.1038/npg.els.0000687 (2001).
Szostak, J. W., Bartel, D. P. & Luisi, P. L. Synthesizing life. Nature 409, 387–390 (2001).
Zuker, M. Mfold web server for nucleic acid folding and hybridization prediction. Nucleic Acids Res. 31, 3406–3415 (2003).
We thank Y. Shi, J. W. Szostak and N. Yan for helpful discussions and comments on the manuscript. We also thank Z. Chen, D. Li, Q. Li, P. Liang, X. Sheng, L. Sun, P. Yin and P. Xu for assistance with the recombinant ASFV pol X purification and isotope-labelling experiments, J. Liu and H. Deng for assistance with the MS experiments and Z. Li and X. Tao for assistance with the preparation of Fig. 1a. This work was supported in part by funding from the National Natural Science Foundation of China (grant no. 31470532, grant no. 91543102 and grant no. 21532004), the Ministry of Science and Technology of China (grant no. 2015CB553402 and grant no. 2013CB932800), the Tsinghua University Initiative Scientific Research Program, the Tsinghua University–Peking University Center for Life Sciences and the Collaborative Innovation Center for Diagnosis and Treatment of Infectious Diseases.
The authors have filed a provisional patent application related to this work.
About this article
Cite this article
Wang, Z., Xu, W., Liu, L. et al. A synthetic molecular system capable of mirror-image genetic replication and transcription. Nature Chem 8, 698–704 (2016). https://doi.org/10.1038/nchem.2517
Chassis engineering for microbial production of chemicals: from natural microbes to synthetic organisms
Current Opinion in Biotechnology (2020)
BMC Biology (2020)
Molecular Physics (2020)
Total synthesis of TRADD death domain with arginine N-GlcNAcylation by hydrazide-based native chemical ligation
Chinese Chemical Letters (2020)