Functional characterization of the meiosis-specific DNA double-strand break inducing factor SPO-11 from C. elegans

Abstract The programmed induction of meiotic DNA double-strand breaks (DSBs) by the evolutionarily conserved SPO-11 protein, which is structurally related to archaeal Topo VIA topoisomerases, triggers meiotic recombination. Identification of several meiosis-specific factors that are required for SPO...

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Autores principales:	Hsin-Yi Yeh, Sheng-Wei Lin, Yi-Chun Wu, Nei-Li Chan, Peter Chi
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2017
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/cac9161265444b8b8f35082c0e583989
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spelling	oai:doaj.org-article:cac9161265444b8b8f35082c0e5839892021-12-02T15:04:53ZFunctional characterization of the meiosis-specific DNA double-strand break inducing factor SPO-11 from C. elegans10.1038/s41598-017-02641-z2045-2322https://doaj.org/article/cac9161265444b8b8f35082c0e5839892017-05-01T00:00:00Zhttps://doi.org/10.1038/s41598-017-02641-zhttps://doaj.org/toc/2045-2322Abstract The programmed induction of meiotic DNA double-strand breaks (DSBs) by the evolutionarily conserved SPO-11 protein, which is structurally related to archaeal Topo VIA topoisomerases, triggers meiotic recombination. Identification of several meiosis-specific factors that are required for SPO-11-mediated DSB formation raises the question whether SPO-11 alone can cleave DNA. Here, we have developed procedures to express and purify C. elegans SPO-11 in a soluble, untagged, and monodispersed form. Our biochemical and biophysical analyses demonstrate that SPO-11 is monomeric and binds DNA, double-stranded DNA in particular. Importantly, SPO-11 does not exhibit DNA cleavage activity under a wide range of reaction conditions, suggesting that co-factors are needed for DSB induction activity. Our SPO-11 purification system and the findings reported herein should facilitate future mechanistic studies directed at delineating the mechanism of action of the SPO-11 ensemble in meiotic DSB formation.Hsin-Yi YehSheng-Wei LinYi-Chun WuNei-Li ChanPeter ChiNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 7, Iss 1, Pp 1-11 (2017)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Hsin-Yi Yeh Sheng-Wei Lin Yi-Chun Wu Nei-Li Chan Peter Chi Functional characterization of the meiosis-specific DNA double-strand break inducing factor SPO-11 from C. elegans
description	Abstract The programmed induction of meiotic DNA double-strand breaks (DSBs) by the evolutionarily conserved SPO-11 protein, which is structurally related to archaeal Topo VIA topoisomerases, triggers meiotic recombination. Identification of several meiosis-specific factors that are required for SPO-11-mediated DSB formation raises the question whether SPO-11 alone can cleave DNA. Here, we have developed procedures to express and purify C. elegans SPO-11 in a soluble, untagged, and monodispersed form. Our biochemical and biophysical analyses demonstrate that SPO-11 is monomeric and binds DNA, double-stranded DNA in particular. Importantly, SPO-11 does not exhibit DNA cleavage activity under a wide range of reaction conditions, suggesting that co-factors are needed for DSB induction activity. Our SPO-11 purification system and the findings reported herein should facilitate future mechanistic studies directed at delineating the mechanism of action of the SPO-11 ensemble in meiotic DSB formation.
format	article
author	Hsin-Yi Yeh Sheng-Wei Lin Yi-Chun Wu Nei-Li Chan Peter Chi
author_facet	Hsin-Yi Yeh Sheng-Wei Lin Yi-Chun Wu Nei-Li Chan Peter Chi
author_sort	Hsin-Yi Yeh
title	Functional characterization of the meiosis-specific DNA double-strand break inducing factor SPO-11 from C. elegans
title_short	Functional characterization of the meiosis-specific DNA double-strand break inducing factor SPO-11 from C. elegans
title_full	Functional characterization of the meiosis-specific DNA double-strand break inducing factor SPO-11 from C. elegans
title_fullStr	Functional characterization of the meiosis-specific DNA double-strand break inducing factor SPO-11 from C. elegans
title_full_unstemmed	Functional characterization of the meiosis-specific DNA double-strand break inducing factor SPO-11 from C. elegans
title_sort	functional characterization of the meiosis-specific dna double-strand break inducing factor spo-11 from c. elegans
publisher	Nature Portfolio
publishDate	2017
url	https://doaj.org/article/cac9161265444b8b8f35082c0e583989
work_keys_str_mv	AT hsinyiyeh functionalcharacterizationofthemeiosisspecificdnadoublestrandbreakinducingfactorspo11fromcelegans AT shengweilin functionalcharacterizationofthemeiosisspecificdnadoublestrandbreakinducingfactorspo11fromcelegans AT yichunwu functionalcharacterizationofthemeiosisspecificdnadoublestrandbreakinducingfactorspo11fromcelegans AT neilichan functionalcharacterizationofthemeiosisspecificdnadoublestrandbreakinducingfactorspo11fromcelegans AT peterchi functionalcharacterizationofthemeiosisspecificdnadoublestrandbreakinducingfactorspo11fromcelegans
_version_	1718389043019907072

Functional characterization of the meiosis-specific DNA double-strand break inducing factor SPO-11 from C. elegans

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