Base pairing and miscoding properties of 1,N(6)-ethenoadenine- and 3,N(4)-ethenocytosine-containing RNA oligonucleotides

Calabretta, Alessandro; Leumann, Christian J. (2013). Base pairing and miscoding properties of 1,N(6)-ethenoadenine- and 3,N(4)-ethenocytosine-containing RNA oligonucleotides. Biochemistry, 52(11), pp. 1990-1997. American Chemical Society 10.1021/bi400116y

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Two RNA phosphoramidites containing the bases 1,N(6)-ethenoadenine (εA) and 3,N(4)-ethenocytosine (εC) were synthesized. These building blocks were incorporated into two 12-mer oligoribonucleotides for evaluation of the base pairing properties of these base lesions by UV melting curve (Tm) and circular dichroism measurements. The Tm data of the resulting duplexes with the etheno modifications opposing all natural bases showed a substantial destabilization compared to the corresponding natural duplexes, confirming their inability to form base pairs. The coding properties of these lesions were further investigated by introducing them into 31-mer oligonucleotides and assessing their ability to serve as templates in primer extension reactions with HIV, AMV, and MMLV reverse transcriptases (RT). Primer extension reactions showed complete arrest of the incorporation process using MMLV RT and AMV RT, while HIV RT preferentially incorporates dAMP opposite εA and dAMP as well as dTMP opposite εC. The properties of these RNA lesions are discussed in the context of its putative biological role.

Item Type:

Journal Article (Original Article)

Division/Institute:

08 Faculty of Science > Departement of Chemistry and Biochemistry

UniBE Contributor:

Calabretta, Alessandro and Leumann, Christian

Subjects:

500 Science > 570 Life sciences; biology
500 Science > 540 Chemistry

ISSN:

0006-2960

Publisher:

American Chemical Society

Funders:

[4] Swiss National Science Foundation
[UNSPECIFIED] Universität Bern

Language:

English

Submitter:

Christian Leumann

Date Deposited:

08 Jan 2014 16:51

Last Modified:

14 Apr 2015 16:18

Publisher DOI:

10.1021/bi400116y

PubMed ID:

23425279

BORIS DOI:

10.7892/boris.39686

URI:

https://boris.unibe.ch/id/eprint/39686

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