| Title: | CRISPR/Cas9-based knockouts reveal that CpRLP1 is a negative regulator of the sex pheromone PR-IP in the Closterium peracerosum-strigosum-littorale complex |
| Author(s): | Kanda N; Ichikawa M; Ono A; Toyoda A; Fujiyama A; Abe J; Tsuchikane Y; Nishiyama T; Sekimoto H; |
| Address: | "Graduate School of Science, Japan Women's University, 2-8-1 Mejirodai, Bunkyo-ku, Tokyo, 112-8681, Japan. Faculty of Science, Japan Women's University, 2-8-1 Mejirodai, Bunkyo-ku, Tokyo, 112-8681, Japan. Center for Information Biology, National Institute of Genetics, 1111 Yata, Mishima, Shizuoka, 411-8540, Japan. Advanced Science Research Center, Kanazawa University, Takaramachi 13-1, Kanazawa, Ishikawa, 920-0934, Japan. Graduate School of Science, Japan Women's University, 2-8-1 Mejirodai, Bunkyo-ku, Tokyo, 112-8681, Japan. sekimoto@fc.jwu.ac.jp. Faculty of Science, Japan Women's University, 2-8-1 Mejirodai, Bunkyo-ku, Tokyo, 112-8681, Japan. sekimoto@fc.jwu.ac.jp" |
| DOI: | 10.1038/s41598-017-18251-8 |
| ISSN/ISBN: | 2045-2322 (Electronic) 2045-2322 (Linking) |
| Abstract: | "Heterothallic strains of the Closterium peracerosum-strigosum-littorale (C. psl.) complex have two sexes, mating-type plus (mt(+)) and mating-type minus (mt(-)). Conjugation between these two sexes is regulated by two sex pheromones, protoplast-release-inducing protein (PR-IP) and PR-IP Inducer, which are produced by mt(+) and mt(-) cells, respectively. PR-IP mediates the release of protoplasts from mt(-) cells during mating. In this study, we examined the mechanism of action of CpRLP1 (receptor-like protein 1), which was previously identified in a cDNA microarray analysis as one of the PR-IP-inducible genes. Using CRISPR/Cas9 technology, we generated CpRLP1 knockout mutants in mt(-) cells of the C. psl. complex. When the knockout mt(-) cells were mixed with wild-type mt(+) cells, conjugation was severely reduced. Many cells released protoplasts without pairing, suggesting a loss of synchronization between the two mating partners. Furthermore, the knockout mutants were hypersensitive to PR-IP. We conclude that CpRLP1 is a negative regulator of PR-IP that regulates the timing of protoplast release in conjugating C. psl. cells. As the first report of successful gene knockout in the class Charophyceae, this study provides a basis for research aimed at understanding the ancestral roles of genes that are indispensable for the development of land plants" |
| Keywords: | Amino Acid Sequence Arabidopsis Proteins/*genetics Biological Phenomena/genetics CRISPR-Cas Systems/genetics/*physiology Carrier Proteins/*genetics Closterium/genetics/*physiology Clustered Regularly Interspaced Short Palindromic Repeats/*genetics/physiol; |
| Notes: | "MedlineKanda, Naho Ichikawa, Machiko Ono, Ayaka Toyoda, Atsushi Fujiyama, Asao Abe, Jun Tsuchikane, Yuki Nishiyama, Tomoaki Sekimoto, Hiroyuki eng Research Support, Non-U.S. Gov't England 2017/12/21 Sci Rep. 2017 Dec 19; 7(1):17873. doi: 10.1038/s41598-017-18251-8" |
