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Compounds that enhance the tailing activity of Moloney murine leukemia virus reverse transcriptase

In a previous study, we showed that MMLV-RT has a strong terminal transferase activity, and that the C-, G-, and T-tailing activities are enhanced by dGMP, dCMP, and dAMP, respectively. In this study, to achieve faster reaction and higher tailing efficiency, we screened other compounds for the abili...

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Autores principales: Ohtsubo, Yoshiyuki, Nagata, Yuji, Tsuda, Masataka
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group UK 2017
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5529421/
https://www.ncbi.nlm.nih.gov/pubmed/28747695
http://dx.doi.org/10.1038/s41598-017-04765-8
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author Ohtsubo, Yoshiyuki
Nagata, Yuji
Tsuda, Masataka
author_facet Ohtsubo, Yoshiyuki
Nagata, Yuji
Tsuda, Masataka
author_sort Ohtsubo, Yoshiyuki
collection PubMed
description In a previous study, we showed that MMLV-RT has a strong terminal transferase activity, and that the C-, G-, and T-tailing activities are enhanced by dGMP, dCMP, and dAMP, respectively. In this study, to achieve faster reaction and higher tailing efficiency, we screened other compounds for the ability to enhance the tailing activities of MMLV-RT, and determined the corresponding optimal concentrations. The C-, G-, and T-tailing activities were enhanced by guanine, cytosine, and adenine, respectively, and by derivatives thereof, suggesting a transient Watson-Click base pairing between an enhancer molecule and the nucleotide to be incorporated. In the presence of some additives (GMP and GDP for C-tailing and CMP for G-tailing), the tail length increased continuously, resulting in tail lengths of 7 to 15 (GMP and GDP) or 13 to 22 (CMP) nucleotides. Among the compounds that do not induce continuous addition, adenosine, deoxycytidine, and deoxyguanosine mostly enhanced T-, G-, and C-tailings, respectively. The enhancing chemicals described here will improve the feasibility of N-tailing by MMLV-RT in various biotechnological applications.
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spelling pubmed-55294212017-08-02 Compounds that enhance the tailing activity of Moloney murine leukemia virus reverse transcriptase Ohtsubo, Yoshiyuki Nagata, Yuji Tsuda, Masataka Sci Rep Article In a previous study, we showed that MMLV-RT has a strong terminal transferase activity, and that the C-, G-, and T-tailing activities are enhanced by dGMP, dCMP, and dAMP, respectively. In this study, to achieve faster reaction and higher tailing efficiency, we screened other compounds for the ability to enhance the tailing activities of MMLV-RT, and determined the corresponding optimal concentrations. The C-, G-, and T-tailing activities were enhanced by guanine, cytosine, and adenine, respectively, and by derivatives thereof, suggesting a transient Watson-Click base pairing between an enhancer molecule and the nucleotide to be incorporated. In the presence of some additives (GMP and GDP for C-tailing and CMP for G-tailing), the tail length increased continuously, resulting in tail lengths of 7 to 15 (GMP and GDP) or 13 to 22 (CMP) nucleotides. Among the compounds that do not induce continuous addition, adenosine, deoxycytidine, and deoxyguanosine mostly enhanced T-, G-, and C-tailings, respectively. The enhancing chemicals described here will improve the feasibility of N-tailing by MMLV-RT in various biotechnological applications. Nature Publishing Group UK 2017-07-26 /pmc/articles/PMC5529421/ /pubmed/28747695 http://dx.doi.org/10.1038/s41598-017-04765-8 Text en © The Author(s) 2017 Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.
spellingShingle Article
Ohtsubo, Yoshiyuki
Nagata, Yuji
Tsuda, Masataka
Compounds that enhance the tailing activity of Moloney murine leukemia virus reverse transcriptase
title Compounds that enhance the tailing activity of Moloney murine leukemia virus reverse transcriptase
title_full Compounds that enhance the tailing activity of Moloney murine leukemia virus reverse transcriptase
title_fullStr Compounds that enhance the tailing activity of Moloney murine leukemia virus reverse transcriptase
title_full_unstemmed Compounds that enhance the tailing activity of Moloney murine leukemia virus reverse transcriptase
title_short Compounds that enhance the tailing activity of Moloney murine leukemia virus reverse transcriptase
title_sort compounds that enhance the tailing activity of moloney murine leukemia virus reverse transcriptase
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5529421/
https://www.ncbi.nlm.nih.gov/pubmed/28747695
http://dx.doi.org/10.1038/s41598-017-04765-8
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