caa a22 4500 605498342 CHVBK 20210128100545.0 cr unu---uuuuu 210128e20150801xx s 000 0 eng 10.1007/s00253-015-6621-2 doi (NATIONALLICENCE)springer-10.1007/s00253-015-6621-2 Mutant screening for oncogenes of Ewing's sarcoma using yeast [Elektronische Daten] [Takao Kitagawa, Hajime Okita, Byron Baron, Kazuhiro Tokuda, Mikiko Nakamura, Yufeng Wang, Junko Akada, Hisashi Hoshida, Rinji Akada, Yasuhiro Kuramitsu, Kazuyuki Nakamura] Many fusion genes, which are the result of chromosomal translocation and work as an oncogene, have been recently identified, but their mode of actions is still unclear. Here, we performed a yeast mutant screening for oncogenes of Ewing's sarcoma to easily identify essential regions responsible for fusion protein functions using a yeast genetic system. Three kinds of oncogenes including EWS/FLI1, EWS/ERG, and EWS/E1AF exhibited growth inhibition in yeast. In this screening, we identified 13 single amino acid substitution mutants which could suppress growth inhibition by oncogenes. All of the point mutation positions of the EWS/ETS family proteins were located within the ETS domain, which is responsible for the interaction with a specific DNA motif. Eight-mutated residues within the ETS domain matched to 13 completely conserved amino acid residues in the human ETS domains. Moreover, mutants also showed reduced transcriptional activities on the DKK2 promoter, which is upregulated by the EWS/ETS family, compared to that of the wild type. These results suggest that the ETS domain in the EWS/ETS family proteins may be a primary target for growth inhibition of Ewing's sarcoma and that this yeast screening system can be applied for the functional screening of the oncogenes. Springer-Verlag Berlin Heidelberg, 2015 Ewing's sarcoma nationallicence EWS/ETS family nationallicence ETS domain nationallicence Yeast nationallicence Kitagawa Takao Department of Biochemistry and Functional Proteomics, Yamaguchi University Graduate School of Medicine, 1-1-1 Minami-Kogushi, 755-8505, Ube, Yamaguchi, Japan aut Okita Hajime Department of Pediatric Hematology and Oncology Research, National Research Institute for Child Health and Development, 2-10-1, Okura, Setagaya-ku, 157-8535, Tokyo, Japan aut Baron Byron Department of Biochemistry and Functional Proteomics, Yamaguchi University Graduate School of Medicine, 1-1-1 Minami-Kogushi, 755-8505, Ube, Yamaguchi, Japan aut Tokuda Kazuhiro Department of Biochemistry and Functional Proteomics, Yamaguchi University Graduate School of Medicine, 1-1-1 Minami-Kogushi, 755-8505, Ube, Yamaguchi, Japan aut Nakamura Mikiko Innovative center, Yamaguchi University, Tokiwadai, 755-8611, Ube, Japan aut Wang Yufeng Department of Biochemistry and Functional Proteomics, Yamaguchi University Graduate School of Medicine, 1-1-1 Minami-Kogushi, 755-8505, Ube, Yamaguchi, Japan aut Akada Junko Department of Biochemistry and Functional Proteomics, Yamaguchi University Graduate School of Medicine, 1-1-1 Minami-Kogushi, 755-8505, Ube, Yamaguchi, Japan aut Hoshida Hisashi Department of Applied Molecular Bioscience, Graduate School of Medicine, Yamaguchi University, Tokiwadai, 755-8611, Ube, Japan aut Akada Rinji Department of Applied Molecular Bioscience, Graduate School of Medicine, Yamaguchi University, Tokiwadai, 755-8611, Ube, Japan aut Kuramitsu Yasuhiro Department of Biochemistry and Functional Proteomics, Yamaguchi University Graduate School of Medicine, 1-1-1 Minami-Kogushi, 755-8505, Ube, Yamaguchi, Japan aut Nakamura Kazuyuki Center of Clinical Laboratories in Tokuyama Medical Association Hospital, 745-0846, Shunan, Japan aut Applied Microbiology and Biotechnology Springer Berlin Heidelberg 99/16(2015-08-01), 6737-6744 0175-7598 99:16<6737 2015 99 253 https://doi.org/10.1007/s00253-015-6621-2 text/html Onlinezugriff via DOI BK010053 XK010053 XK010000 Metadata rights reserved Springer special CC-BY-NC licence nationallicence 1 research-article jats NATIONALLICENCE NATIONALLICENCE NL-springer NATIONALLICENCE 856 40 https://doi.org/10.1007/s00253-015-6621-2 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Kitagawa Takao Department of Biochemistry and Functional Proteomics, Yamaguchi University Graduate School of Medicine, 1-1-1 Minami-Kogushi, 755-8505, Ube, Yamaguchi, Japan aut NATIONALLICENCE 700 1- Okita Hajime Department of Pediatric Hematology and Oncology Research, National Research Institute for Child Health and Development, 2-10-1, Okura, Setagaya-ku, 157-8535, Tokyo, Japan aut NATIONALLICENCE 700 1- Baron Byron Department of Biochemistry and Functional Proteomics, Yamaguchi University Graduate School of Medicine, 1-1-1 Minami-Kogushi, 755-8505, Ube, Yamaguchi, Japan aut NATIONALLICENCE 700 1- Tokuda Kazuhiro Department of Biochemistry and Functional Proteomics, Yamaguchi University Graduate School of Medicine, 1-1-1 Minami-Kogushi, 755-8505, Ube, Yamaguchi, Japan aut NATIONALLICENCE 700 1- Nakamura Mikiko Innovative center, Yamaguchi University, Tokiwadai, 755-8611, Ube, Japan aut NATIONALLICENCE 700 1- Wang Yufeng Department of Biochemistry and Functional Proteomics, Yamaguchi University Graduate School of Medicine, 1-1-1 Minami-Kogushi, 755-8505, Ube, Yamaguchi, Japan aut NATIONALLICENCE 700 1- Akada Junko Department of Biochemistry and Functional Proteomics, Yamaguchi University Graduate School of Medicine, 1-1-1 Minami-Kogushi, 755-8505, Ube, Yamaguchi, Japan aut NATIONALLICENCE 700 1- Hoshida Hisashi Department of Applied Molecular Bioscience, Graduate School of Medicine, Yamaguchi University, Tokiwadai, 755-8611, Ube, Japan aut NATIONALLICENCE 700 1- Akada Rinji Department of Applied Molecular Bioscience, Graduate School of Medicine, Yamaguchi University, Tokiwadai, 755-8611, Ube, Japan aut NATIONALLICENCE 700 1- Kuramitsu Yasuhiro Department of Biochemistry and Functional Proteomics, Yamaguchi University Graduate School of Medicine, 1-1-1 Minami-Kogushi, 755-8505, Ube, Yamaguchi, Japan aut NATIONALLICENCE 700 1- Nakamura Kazuyuki Center of Clinical Laboratories in Tokuyama Medical Association Hospital, 745-0846, Shunan, Japan aut NATIONALLICENCE 773 0- Applied Microbiology and Biotechnology Springer Berlin Heidelberg 99/16(2015-08-01), 6737-6744 0175-7598 99:16<6737 2015 99 253