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

Abstract 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...

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Autores principales:	Yoshiyuki Ohtsubo, Yuji Nagata, Masataka Tsuda
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2017
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/f5ec3428e7b04468bb18e47b744af692
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spelling	oai:doaj.org-article:f5ec3428e7b04468bb18e47b744af6922021-12-02T16:06:52ZCompounds that enhance the tailing activity of Moloney murine leukemia virus reverse transcriptase10.1038/s41598-017-04765-82045-2322https://doaj.org/article/f5ec3428e7b04468bb18e47b744af6922017-07-01T00:00:00Zhttps://doi.org/10.1038/s41598-017-04765-8https://doaj.org/toc/2045-2322Abstract 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.Yoshiyuki OhtsuboYuji NagataMasataka TsudaNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 7, Iss 1, Pp 1-6 (2017)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Yoshiyuki Ohtsubo Yuji Nagata Masataka Tsuda Compounds that enhance the tailing activity of Moloney murine leukemia virus reverse transcriptase
description	Abstract 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.
format	article
author	Yoshiyuki Ohtsubo Yuji Nagata Masataka Tsuda
author_facet	Yoshiyuki Ohtsubo Yuji Nagata Masataka Tsuda
author_sort	Yoshiyuki Ohtsubo
title	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_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_sort	compounds that enhance the tailing activity of moloney murine leukemia virus reverse transcriptase
publisher	Nature Portfolio
publishDate	2017
url	https://doaj.org/article/f5ec3428e7b04468bb18e47b744af692
work_keys_str_mv	AT yoshiyukiohtsubo compoundsthatenhancethetailingactivityofmoloneymurineleukemiavirusreversetranscriptase AT yujinagata compoundsthatenhancethetailingactivityofmoloneymurineleukemiavirusreversetranscriptase AT masatakatsuda compoundsthatenhancethetailingactivityofmoloneymurineleukemiavirusreversetranscriptase
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Compounds that enhance the tailing activity of Moloney murine leukemia virus reverse transcriptase

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