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Filament formation and robust strand exchange activities of the rice DMC1A and DMC1B proteins

The DMC1 protein, a meiosis-specific DNA recombinase, catalyzes strand exchange between homologous chromosomes. In rice, two Dmc1 genes, Dmc1A and Dmc1B, have been reported. Although the Oryza sativa DMC1A protein has been partially characterized, however the biochemical properties of the DMC1B prot...

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Autores principales: Sakane, Isao, Kamataki, Chiaki, Takizawa, Yoshimasa, Nakashima, Marina, Toki, Seiichi, Ichikawa, Hiroaki, Ikawa, Shukuko, Shibata, Takehiko, Kurumizaka, Hitoshi
Formato: Texto
Lenguaje:English
Publicado: Oxford University Press 2008
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2490746/
https://www.ncbi.nlm.nih.gov/pubmed/18583359
http://dx.doi.org/10.1093/nar/gkn405
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author Sakane, Isao
Kamataki, Chiaki
Takizawa, Yoshimasa
Nakashima, Marina
Toki, Seiichi
Ichikawa, Hiroaki
Ikawa, Shukuko
Shibata, Takehiko
Kurumizaka, Hitoshi
author_facet Sakane, Isao
Kamataki, Chiaki
Takizawa, Yoshimasa
Nakashima, Marina
Toki, Seiichi
Ichikawa, Hiroaki
Ikawa, Shukuko
Shibata, Takehiko
Kurumizaka, Hitoshi
author_sort Sakane, Isao
collection PubMed
description The DMC1 protein, a meiosis-specific DNA recombinase, catalyzes strand exchange between homologous chromosomes. In rice, two Dmc1 genes, Dmc1A and Dmc1B, have been reported. Although the Oryza sativa DMC1A protein has been partially characterized, however the biochemical properties of the DMC1B protein have not been defined. In the present study, we expressed the Oryza sativa DMC1A and DMC1B proteins in bacteria and purified them. The purified DMC1A and DMC1B proteins formed helical filaments along single-stranded DNA (ssDNA) and double-stranded DNA (dsDNA), and promoted robust strand exchange between ssDNA and dsDNA over five thousand base pairs in the presence of RPA, as a co-factor. The DMC1A and DMC1B proteins also promoted strand exchange in the absence of RPA with long DNA substrates containing several thousand base pairs. In contrast, the human DMC1 protein strictly required RPA to promote strand exchange with these long DNA substrates. The strand-exchange activity of the Oryza sativa DMC1A protein was much higher than that of the DMC1B protein. Consistently, the DNA-binding activity of the DMC1A protein was higher than that of the DMC1B protein. These biochemical differences between the DMC1A and DMC1B proteins may provide important insight into their functional differences during meiosis in rice.
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spelling pubmed-24907462008-08-01 Filament formation and robust strand exchange activities of the rice DMC1A and DMC1B proteins Sakane, Isao Kamataki, Chiaki Takizawa, Yoshimasa Nakashima, Marina Toki, Seiichi Ichikawa, Hiroaki Ikawa, Shukuko Shibata, Takehiko Kurumizaka, Hitoshi Nucleic Acids Res Molecular Biology The DMC1 protein, a meiosis-specific DNA recombinase, catalyzes strand exchange between homologous chromosomes. In rice, two Dmc1 genes, Dmc1A and Dmc1B, have been reported. Although the Oryza sativa DMC1A protein has been partially characterized, however the biochemical properties of the DMC1B protein have not been defined. In the present study, we expressed the Oryza sativa DMC1A and DMC1B proteins in bacteria and purified them. The purified DMC1A and DMC1B proteins formed helical filaments along single-stranded DNA (ssDNA) and double-stranded DNA (dsDNA), and promoted robust strand exchange between ssDNA and dsDNA over five thousand base pairs in the presence of RPA, as a co-factor. The DMC1A and DMC1B proteins also promoted strand exchange in the absence of RPA with long DNA substrates containing several thousand base pairs. In contrast, the human DMC1 protein strictly required RPA to promote strand exchange with these long DNA substrates. The strand-exchange activity of the Oryza sativa DMC1A protein was much higher than that of the DMC1B protein. Consistently, the DNA-binding activity of the DMC1A protein was higher than that of the DMC1B protein. These biochemical differences between the DMC1A and DMC1B proteins may provide important insight into their functional differences during meiosis in rice. Oxford University Press 2008-08 2008-06-26 /pmc/articles/PMC2490746/ /pubmed/18583359 http://dx.doi.org/10.1093/nar/gkn405 Text en © 2008 The Author(s) http://creativecommons.org/licenses/by-nc/2.0/uk/ This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/2.0/uk/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Molecular Biology
Sakane, Isao
Kamataki, Chiaki
Takizawa, Yoshimasa
Nakashima, Marina
Toki, Seiichi
Ichikawa, Hiroaki
Ikawa, Shukuko
Shibata, Takehiko
Kurumizaka, Hitoshi
Filament formation and robust strand exchange activities of the rice DMC1A and DMC1B proteins
title Filament formation and robust strand exchange activities of the rice DMC1A and DMC1B proteins
title_full Filament formation and robust strand exchange activities of the rice DMC1A and DMC1B proteins
title_fullStr Filament formation and robust strand exchange activities of the rice DMC1A and DMC1B proteins
title_full_unstemmed Filament formation and robust strand exchange activities of the rice DMC1A and DMC1B proteins
title_short Filament formation and robust strand exchange activities of the rice DMC1A and DMC1B proteins
title_sort filament formation and robust strand exchange activities of the rice dmc1a and dmc1b proteins
topic Molecular Biology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2490746/
https://www.ncbi.nlm.nih.gov/pubmed/18583359
http://dx.doi.org/10.1093/nar/gkn405
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