Effect of terminal 3'-hydroxymethyl modification of an RNA primer on nonenzymatic primer extension

Ayan Pal; Rajat S Das; Weicheng Zhang; Megan Lang; Larry W McLaughlin; Jack W Szostak

doi:10.1039/c6cc06925h

Effect of terminal 3'-hydroxymethyl modification of an RNA primer on nonenzymatic primer extension

Chem Commun (Camb). 2016 Sep 29;52(80):11905-11907. doi: 10.1039/c6cc06925h.

Authors

Ayan Pal¹, Rajat S Das², Weicheng Zhang³, Megan Lang⁴, Larry W McLaughlin⁴, Jack W Szostak⁵

Affiliations

¹ Howard Hughes Medical Institute, Department of Molecular Biology and Center for Computational and Integrative Biology, Massachusetts General Hospital, 185 Cambridge Street, Boston, Massachusetts 02114, USA. szostak@molbio.mgh.harvard.edu and Department of Genetics, Harvard Medical School, 77 Avenue Louis Pasteur, Boston, Massachusetts 02115, USA.
² Boston College, Department of Chemistry, Merkert Chemistry Center, 2609 Beacon Street, Chestnut Hill, Massachusetts 02467, USA and Alnylam Pharmaceuticals, 300 Third Street, Cambridge, Massachusetts 02142, USA.
³ Howard Hughes Medical Institute, Department of Molecular Biology and Center for Computational and Integrative Biology, Massachusetts General Hospital, 185 Cambridge Street, Boston, Massachusetts 02114, USA. szostak@molbio.mgh.harvard.edu and Department of Chemistry and Chemical Biology, Harvard University, 12 Oxford St., Cambridge, Massachusetts 02138, USA.
⁴ Boston College, Department of Chemistry, Merkert Chemistry Center, 2609 Beacon Street, Chestnut Hill, Massachusetts 02467, USA.
⁵ Howard Hughes Medical Institute, Department of Molecular Biology and Center for Computational and Integrative Biology, Massachusetts General Hospital, 185 Cambridge Street, Boston, Massachusetts 02114, USA. szostak@molbio.mgh.harvard.edu and Department of Genetics, Harvard Medical School, 77 Avenue Louis Pasteur, Boston, Massachusetts 02115, USA and Department of Chemistry and Chemical Biology, Harvard University, 12 Oxford St., Cambridge, Massachusetts 02138, USA.

PMID: 27722518
DOI: 10.1039/c6cc06925h

Abstract

The significance of the precise position of the hydroxyl at the 3'-end of an RNA primer for nonenzymatic template-directed primer extension is not well understood. We show that an RNA primer terminating in 3'-hydroxymethyl-2',3'-dideoxy-guanosine has greatly diminished activity, suggesting that the spatial preorganization of the terminal sugar contributes significantly to the efficiency of primer extension.

MeSH terms

Chromatography, High Pressure Liquid
Electrophoresis, Gel, Pulsed-Field
Guanosine / chemistry
Mass Spectrometry
Nucleic Acid Amplification Techniques / methods*
RNA / analysis
RNA / chemical synthesis
RNA / chemistry*
Stereoisomerism

Substances

RNA primers
Guanosine
RNA