The double-stranded break-forming activity of plant SPO11s and a novel rice SPO11 revealed by a Drosophila bioassay

• Published in: BMC molecular biology Vol. 13; no. 1; p. 1
• Main Authors: Shingu, Yoshinori, Tokai, Takeshi, Agawa, Yasuo, Toyota, Kentaro, Ahamed, Selina, Kawagishi-Kobayashi, Makiko, Komatsu, Akira, Mikawa, Tsutomu, Yamamoto, Masa-Toshi, Wakasa, Kyo, Shibata, Takehiko, Kusano, Kohji
• Format: Journal Article
• Language: English
• Published: England BioMed Central Ltd 16.01.2012; BioMed Central
• Subjects: Amino Acid Sequence; Animals; Anthers; Arabidopsis; Arabidopsis - metabolism; Arabidopsis Proteins - genetics; Arabidopsis Proteins - metabolism; Biological Assay; Chromatin; Chromosomes; Deoxyribonucleic acid; DNA; DNA Breaks, Double-Stranded; DNA damage; DNA Topoisomerases - genetics; DNA Topoisomerases - metabolism; Drosophila; Drosophila - growth & development; Endodeoxyribonucleases - genetics; Endodeoxyribonucleases - metabolism; Genes; Genetic analysis; Genetic recombination; Meiosis; Molecular Sequence Data; Oocytes - metabolism; Oryza - metabolism; Oryza sativa; Physiological aspects; Plant Proteins - genetics; Plant Proteins - metabolism; Pollen; Proteins; Recombination; RNA, Messenger - metabolism; Transgenes; Yeast
• ISSN: 1471-2199; 1471-2199
• DOI: 10.1186/1471-2199-13-1

SPO11 is a key protein for promoting meiotic recombination, by generating chromatin locus- and timing-specific DNA double-strand breaks (DSBs). The DSB activity of SPO11 was shown by genetic analyses, but whether SPO11 exerts DSB-forming activity by itself is still an unanswered question. DSB formation by SPO11 has not been detected by biochemical means, probably because of a lack of proper protein-folding, posttranslational modifications, and/or specific SPO11-interacting proteins required for this activity. In addition, plants have multiple SPO11-homologues. To determine whether SPO11 can cleave DNA by itself, and to identify which plant SPO11 homologue cleaves DNA, we developed a Drosophila bioassay system that detects the DSB signals generated by a plant SPO11 homologue expressed ectopically. We cytologically and genetically demonstrated the DSB activities of Arabidopsis AtSPO11-1 and AtSPO11-2, which are required for meiosis, in the absence of other plant proteins. Using this bioassay, we further found that a novel SPO11-homologue, OsSPO11D, which has no counterpart in Arabidopsis, displays prominent DSB-forming activity. Quantitative analyses of the rice SPO11 transcripts revealed the specific increase in OsSPO11D mRNA in the anthers containing meiotic pollen mother cells. The Drosophila bioassay system successfully demonstrated that some plant SPO11 orthologues have intrinsic DSB activities. Furthermore, we identified a novel SPO11 homologue, OsSPO11D, with robust DSB activity and a possible meiotic function.