Efficient and rapid template-directed nucleic acid copying using 2'-amino-2',3'-dideoxyribonucleoside-5'-phosphorimidazolide monomers
| Autor: | J. Craig Blain, Jack W. Szostak, Jason P. Schrum, Mathangi Krishnamurthy, Alonso Ricardo |
|---|---|
| Rok vydání: | 2009 |
| Předmět: |
Protocell
010402 general chemistry 01 natural sciences Biochemistry Catalysis Article 03 medical and health sciences chemistry.chemical_compound Colloid and Surface Chemistry Nucleic Acids Nucleotide 030304 developmental biology chemistry.chemical_classification 0303 health sciences Imidazoles RNA General Chemistry DNA Templates Genetic Combinatorial chemistry Dideoxynucleosides 0104 chemical sciences Monomer Template chemistry Polynucleotide Spectrometry Mass Matrix-Assisted Laser Desorption-Ionization Nucleic acid |
| Zdroj: | Journal of the American Chemical Society |
| ISSN: | 1520-5126 |
| Popis: | The development of a sequence-general nucleic acid copying system is an essential step in the assembly of a synthetic protocell, an autonomously replicating spatially localized chemical system capable of spontaneous Darwinian evolution. Previously described nonenzymatic template-copying experiments have validated the concept of nonenzymatic replication, but have not yet achieved robust, sequence-general polynucleotide replication. The 5'-phosphorimidazolides of the 2'-amino-2',3'-dideoxyribonucleotides are attractive as potential monomers for such a system because they polymerize by forming 2'--5' linkages, which are favored in nonenzymatic polymerization reactions using similarly activated ribonucleotides on RNA templates. Furthermore, the 5'-activated 2'-amino nucleotides do not cyclize. We recently described the rapid and efficient nonenzymatic copying of a DNA homopolymer template (dC(15)) encapsulated within fatty acid vesicles using 2'-amino-2',3'-dideoxyguanosine-5'-phosphorimidazolide as the activated monomer. However, to realize a true Darwinian system, the template-copying chemistry must be able to copy most sequences and their complements to allow for the transmission of information from generation to generation. Here, we describe the copying of a series of nucleic acid templates using 2'-amino-2',3'-dideoxynucleotide-5'-phosphorimidazolides. Polymerization reactions proceed rapidly to completion on short homopolymer RNA and LNA templates, which favor an A-type duplex geometry. We show that more efficiently copied sequences are generated by replacing the adenine nucleobase with diaminopurine, and uracil with C5-(1-propynyl)uracil. Finally, we explore the copying of longer, mixed-sequence RNA templates to assess the sequence-general copying ability of 2'-amino-2',3'-dideoxynucleoside-5'-phosphorimidazolides. Our results are a significant step forward in the realization of a self-replicating genetic polymer compatible with protocell template copying and suggest that N2'--P5'-phosphoramidate DNA may have the potential to function as a self-replicating system. |
| Databáze: | OpenAIRE |
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