Synthesis of phosphoramidate-linked DNA by a modified DNA polymerase.


Journal

Proceedings of the National Academy of Sciences of the United States of America
ISSN: 1091-6490
Titre abrégé: Proc Natl Acad Sci U S A
Pays: United States
ID NLM: 7505876

Informations de publication

Date de publication:
31 03 2020
Historique:
pubmed: 20 3 2020
medline: 15 8 2020
entrez: 20 3 2020
Statut: ppublish

Résumé

All known polymerases copy genetic material by catalyzing phosphodiester bond formation. This highly conserved activity proceeds by a common mechanism, such that incorporated nucleoside analogs terminate chain elongation if the resulting primer strand lacks a terminal hydroxyl group. Even conservatively substituted 3'-amino nucleotides generally act as chain terminators, and no enzymatic pathway for their polymerization has yet been found. Although 3'-amino nucleotides can be chemically coupled to yield stable oligonucleotides containing N3'→P5' phosphoramidate (NP) bonds, no such internucleotide linkages are known to occur in nature. Here, we report that 3'-amino terminated primers are, in fact, slowly extended by the DNA polymerase from

Identifiants

pubmed: 32188786
pii: 1922400117
doi: 10.1073/pnas.1922400117
pmc: PMC7132125
doi:

Substances chimiques

Amides 0
Oligonucleotides 0
Phosphoric Acids 0
RNA 63231-63-0
DNA 9007-49-2
phosphoramidic acid 9Q189608GB
DNA-Directed DNA Polymerase EC 2.7.7.7

Banques de données

PDB
['6UR2', '6UR4', '6UR9', '6US5']

Types de publication

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

Langues

eng

Sous-ensembles de citation

IM

Pagination

7276-7283

Subventions

Organisme : Howard Hughes Medical Institute
Pays : United States

Informations de copyright

Copyright © 2020 the Author(s). Published by PNAS.

Déclaration de conflit d'intérêts

Competing interest statement: The authors have filed for a patent related to this work.

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Auteurs

Victor S Lelyveld (VS)

Department of Molecular Biology, Center for Computational and Integrative Biology, Massachusetts General Hospital, Boston, MA 02114.
Department of Genetics, Harvard Medical School, Boston, MA 02115.

Wen Zhang (W)

Department of Molecular Biology, Center for Computational and Integrative Biology, Massachusetts General Hospital, Boston, MA 02114.
Department of Genetics, Harvard Medical School, Boston, MA 02115.

Jack W Szostak (JW)

Department of Molecular Biology, Center for Computational and Integrative Biology, Massachusetts General Hospital, Boston, MA 02114; szostak@molbio.mgh.harvard.edu.
Department of Genetics, Harvard Medical School, Boston, MA 02115.
Department of Chemistry and Chemical Biology, Harvard University, Cambridge, MA 02138.
Howard Hughes Medical Institute, Massachusetts General Hospital, Boston, MA 02114.

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Classifications MeSH