title: Downstream Oligonucleotides Strongly Enhance the Affinity of GMP to RNA Primer-Template Complexes.
abstract: Origins of life hypotheses often invoke a transitional phase of nonenzymatic template-directed RNA replication prior to the emergence of ribozyme-catalyzed copying of genetic information. Here, using NMR and ITC, we interrogate the binding affinity of guanosine 5'-monophosphate (GMP) for primer-template complexes when either another GMP, or a helper oligonucleotide, can bind downstream. Binding of GMP to a primer-template complex cannot be significantly enhanced by the possibility of downstream monomer binding, because the affinity of the downstream monomer is weaker than that of the first monomer. Strikingly, GMP binding affinity can be enhanced by ca. 2 orders of magnitude when a helper oligonucleotide is stably bound downstream of the monomer binding site. We compare these thermodynamic parameters to those previously reported for T7 RNA polymerase-mediated replication to help address questions of binding affinity in related nonenzymatic processes.
authors.affiliation: Howard Hughes Medical Institute, Department of Molecular Biology and Center for Computational and Integrative Biology, Massachusetts General Hospital , 185 Cambridge Street, Boston, Massachusetts 02114, United States.|Department of Chemistry and Chemical Biology, Harvard University , 12 Oxford Street, Cambridge, Massachusetts 02138, United States.
authors.forename: Chun Pong
authors.identifier: 0000-0001-6381-9011
authors.initials: CP
authors.lastname: Tam
authors.affiliation: Howard Hughes Medical Institute, Department of Molecular Biology and Center for Computational and Integrative Biology, Massachusetts General Hospital , 185 Cambridge Street, Boston, Massachusetts 02114, United States.|Earth Life Science Institute, Tokyo Institute of Technology , 2-12-1-IE-1 Ookayama, Meguro-ku, Tokyo, 152-8550, Japan.
authors.forename: Albert C
authors.initials: AC
authors.lastname: Fahrenbach
authors.affiliation: Howard Hughes Medical Institute, Department of Molecular Biology and Center for Computational and Integrative Biology, Massachusetts General Hospital , 185 Cambridge Street, Boston, Massachusetts 02114, United States.|Department of Biosciences, Åbo Akademi University , Åbo FI-20520, Finland.
authors.forename: Anders
authors.initials: A
authors.lastname: Björkbom
authors.affiliation: Howard Hughes Medical Institute, Department of Molecular Biology and Center for Computational and Integrative Biology, Massachusetts General Hospital , 185 Cambridge Street, Boston, Massachusetts 02114, United States.|Department of Chemistry and Chemical Biology, Harvard University , 12 Oxford Street, Cambridge, Massachusetts 02138, United States.
authors.forename: Noam
authors.initials: N
authors.lastname: Prywes
authors.affiliation: Howard Hughes Medical Institute, Department of Molecular Biology and Center for Computational and Integrative Biology, Massachusetts General Hospital , 185 Cambridge Street, Boston, Massachusetts 02114, United States.
authors.forename: Enver Cagri
authors.identifier: 0000-0001-6673-3635
authors.initials: EC
authors.lastname: Izgu
authors.affiliation: Howard Hughes Medical Institute, Department of Molecular Biology and Center for Computational and Integrative Biology, Massachusetts General Hospital , 185 Cambridge Street, Boston, Massachusetts 02114, United States.|Department of Chemistry and Chemical Biology, Harvard University , 12 Oxford Street, Cambridge, Massachusetts 02138, United States.|Earth Life Science Institute, Tokyo Institute of Technology , 2-12-1-IE-1 Ookayama, Meguro-ku, Tokyo, 152-8550, Japan.
authors.forename: Jack W
authors.initials: JW
authors.lastname: Szostak
mesh_terms: D001665:Binding Sites; D012321:DNA-Directed RNA Polymerases / Q000737:chemistry / Q000378:metabolism; D006157:Guanosine Monophosphate / Q000737:chemistry* / Q000378:metabolism; D009841:Oligonucleotides / Q000737:chemistry* / Q000378:metabolism; D012313:RNA / Q000737:chemistry*; D013816:Thermodynamics; D014764:Viral Proteins / Q000737:chemistry / Q000378:metabolism
chemical_list: D009841:Oligonucleotides; C034477:RNA primers; D014764:Viral Proteins; D012313:RNA; D006157:Guanosine Monophosphate; C062210:bacteriophage T7 RNA polymerase; D012321:DNA-Directed RNA Polymerases