caa a22 4500 467895244 CHVBK 20180406152801.0 cr unu---uuuuu 170328e20060901xx s 000 0 eng 10.1007/s11103-006-9009-y doi (NATIONALLICENCE)springer-10.1007/s11103-006-9009-y Degenerated recognition property of a mitochondrial homing enzyme in the unicellular green alga Chlamydomonas smithii [Elektronische Daten] [Sayuri Kurokawa, Tomohito Yamasaki, Teruaki Komatsu, Kazuo Watanabe, Takeshi Ohama] Target sequence cleavage is the essential step for intron invasion into an intronless allele. DNA cleavage at a specific site is performed by an endonuclease, termed a homing enzyme, which is encoded by an open reading frame within the intron. The recognition properties of them have only been analyzed in vitro, using purified, recombinant homing enzyme and various mutated DNA substrates, but it is unclear whether the homing enzyme behaves similarly in vivo. To answer this question, we determined the recognition properties of I-CsmI in vivo. I-CsmI is a homing enzyme encoded by the open reading frame of the alpha-group I-intron, located in the mitochondrial apocytochrome b gene of the green alga Chlamydomonas smithii. The in vivo recognition properties of it were determined as the frequency of intron invasion into a mutated target site. For this purpose, we utilized hybrid diploid cells developed by crossing alpha-intron-plus C. smithii to intron-minus C. reinhardtii containing mutated target sequences. The intron invasion frequency was much higher than the expected from the in vitro cleavage frequency of the respective mutated substrates. Even the substrates that had very little cleavage in the in vitro experiment were efficiently invaded in vivo, and were accompanied by a large degree of coconversion. Considering the ease of the homing enzyme invading into various mutated target sequences, we propose that the principle bottleneck for lateral intron transmission is not the sequence specificity of the homing enzyme, but instead is limited by the rare occurrence of inter-specific cell fusion. Springer Science+Business Media B.V., 2006 Selfish genetic element nationallicence Group I intron nationallicence Coconversion nationallicence Horizontal transmission nationallicence Adapted recognition nationallicence Kurokawa Sayuri Graduate School of Engineering, Department of Environmental Systems Engineering, Kochi University of Technology (KUT), 782-8502, Tosayamada, Kochi, Japan aut Yamasaki Tomohito Graduate School of Engineering, Department of Environmental Systems Engineering, Kochi University of Technology (KUT), 782-8502, Tosayamada, Kochi, Japan aut Komatsu Teruaki Graduate School of Engineering, Department of Environmental Systems Engineering, Kochi University of Technology (KUT), 782-8502, Tosayamada, Kochi, Japan aut Watanabe Kazuo Institute for Cellular and Molecular Biology, The University of Texas at Austin, Moffett Molecular Biology Bldg, A4800, 2500 Speedway, 78712-1095, Austin, TX, USA aut Ohama Takeshi Graduate School of Engineering, Department of Environmental Systems Engineering, Kochi University of Technology (KUT), 782-8502, Tosayamada, Kochi, Japan aut Plant Molecular Biology Kluwer Academic Publishers 62/1-2(2006-09-01), 141-150 0167-4412 62:1-2<141 2006 62 11103 https://doi.org/10.1007/s11103-006-9009-y text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1007/s11103-006-9009-y text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Kurokawa Sayuri Graduate School of Engineering, Department of Environmental Systems Engineering, Kochi University of Technology (KUT), 782-8502, Tosayamada, Kochi, Japan aut NATIONALLICENCE 700 1- Yamasaki Tomohito Graduate School of Engineering, Department of Environmental Systems Engineering, Kochi University of Technology (KUT), 782-8502, Tosayamada, Kochi, Japan aut NATIONALLICENCE 700 1- Komatsu Teruaki Graduate School of Engineering, Department of Environmental Systems Engineering, Kochi University of Technology (KUT), 782-8502, Tosayamada, Kochi, Japan aut NATIONALLICENCE 700 1- Watanabe Kazuo Institute for Cellular and Molecular Biology, The University of Texas at Austin, Moffett Molecular Biology Bldg, A4800, 2500 Speedway, 78712-1095, Austin, TX, USA aut NATIONALLICENCE 700 1- Ohama Takeshi Graduate School of Engineering, Department of Environmental Systems Engineering, Kochi University of Technology (KUT), 782-8502, Tosayamada, Kochi, Japan aut NATIONALLICENCE 773 0- Plant Molecular Biology Kluwer Academic Publishers 62/1-2(2006-09-01), 141-150 0167-4412 62:1-2<141 2006 62 11103 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer