caa a22 4500 605503648 CHVBK 20210128100611.0 cr unu---uuuuu 210128e20150201xx s 000 0 eng 10.1007/s00253-014-6185-6 doi (NATIONALLICENCE)springer-10.1007/s00253-014-6185-6 Mycoplasma gallisepticum MGA_0676 is a membrane-associated cytotoxic nuclease with a staphylococcal nuclease region essential for nuclear translocation and apoptosis induction in chicken cells [Elektronische Daten] [Jian Xu, Da Teng, Fei Jiang, Yuewei Zhang, Saeed El-Ashram, Hui Wang, Zhenhong Sun, Jinyan He, Junjun Shen, Wenxue Wu, Jinxiang Li] Mycoplasma gallisepticum can infect a wide variety of birds including the commercial poultry. M. gallisepticum MGA_0676 is a putative lipoprotein, which is similar to bacterial thermostable nucleases. But the possible pathogenic effect of M. gallisepticum MGA_0676 has not been investigated so far. In the present study, we cloned the MGA_0676 gene after deletion of the amino-terminal signal sequence and mutagenesis of the Mycoplasma TGA tryptophan codons to TGG and expressed recombinant MGA_0676 protein in Escherichia coli. We identified and characterized MGA_0676 as a Ca2+-dependent cytotoxic nuclease of M. gallisepticum with a staphylococcal nuclease (SNc) region that displays the hallmarks of nucleases. Membrane protein immunoblot analysis and immunogold electron microscopy revealed that MGA_0676 locates on the membrane surface of M. gallisepticum. Furthermore, apoptosis assay using annexin V-FITC and propidium iodide (annexin V/PI) indicated that MGA_0676 played significant roles in apoptosis induction and pathological damages in chicken cells. Moreover, confocal microscopy showed that MGA_0676 localizes in the nuclei of host cells. Besides, after the SNc region was deleted, MGA_0676 lost its ability of nuclear localization, nuclease activity, and cytotoxicity, which revealed that the SNc region is essential for nuclear translocation and induction of apoptosis in chicken cells. The above results suggest that MGA_0676 is an important virulence factor in cellular pathology and may play a unique role in the life cycle events of M. gallisepticum. Springer-Verlag Berlin Heidelberg, 2014 Mycoplasma gallisepticum nationallicence MGA_0676 nationallicence SNc nationallicence Cytotoxic nuclease nationallicence Nuclear translocation nationallicence Apoptosis nationallicence Xu Jian Laboratory of Rapid Diagnostic Technology for Animal Disease, College of Veterinary Medicine, China Agricultural University, 100193, Beijing, People's Republic of China aut Teng Da Laboratory of Rapid Diagnostic Technology for Animal Disease, College of Veterinary Medicine, China Agricultural University, 100193, Beijing, People's Republic of China aut Jiang Fei Laboratory of Rapid Diagnostic Technology for Animal Disease, College of Veterinary Medicine, China Agricultural University, 100193, Beijing, People's Republic of China aut Zhang Yuewei Laboratory of Rapid Diagnostic Technology for Animal Disease, College of Veterinary Medicine, China Agricultural University, 100193, Beijing, People's Republic of China aut El-Ashram Saeed Key Laboratory of Animal Epidemiology and Zoonosis, Ministry of Agriculture, College of Veterinary Medicine, China Agricultural University, 100193, Beijing, People's Republic of China aut Wang Hui Key Laboratory of Animal Epidemiology and Zoonosis, Ministry of Agriculture, College of Veterinary Medicine, China Agricultural University, 100193, Beijing, People's Republic of China aut Sun Zhenhong Taishan Medical University, 271000, Taian, Shandong, People's Republic of China aut He Jinyan Laboratory of Rapid Diagnostic Technology for Animal Disease, College of Veterinary Medicine, China Agricultural University, 100193, Beijing, People's Republic of China aut Shen Junjun Laboratory of Rapid Diagnostic Technology for Animal Disease, College of Veterinary Medicine, China Agricultural University, 100193, Beijing, People's Republic of China aut Wu Wenxue Laboratory of Rapid Diagnostic Technology for Animal Disease, College of Veterinary Medicine, China Agricultural University, 100193, Beijing, People's Republic of China aut Li Jinxiang Chinese Academy of Agricultural Sciences, 100081, Beijing, People's Republic of China aut Applied Microbiology and Biotechnology Springer Berlin Heidelberg 99/4(2015-02-01), 1859-1871 0175-7598 99:4<1859 2015 99 253 https://doi.org/10.1007/s00253-014-6185-6 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/s00253-014-6185-6 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Xu Jian Laboratory of Rapid Diagnostic Technology for Animal Disease, College of Veterinary Medicine, China Agricultural University, 100193, Beijing, People's Republic of China aut NATIONALLICENCE 700 1- Teng Da Laboratory of Rapid Diagnostic Technology for Animal Disease, College of Veterinary Medicine, China Agricultural University, 100193, Beijing, People's Republic of China aut NATIONALLICENCE 700 1- Jiang Fei Laboratory of Rapid Diagnostic Technology for Animal Disease, College of Veterinary Medicine, China Agricultural University, 100193, Beijing, People's Republic of China aut NATIONALLICENCE 700 1- Zhang Yuewei Laboratory of Rapid Diagnostic Technology for Animal Disease, College of Veterinary Medicine, China Agricultural University, 100193, Beijing, People's Republic of China aut NATIONALLICENCE 700 1- El-Ashram Saeed Key Laboratory of Animal Epidemiology and Zoonosis, Ministry of Agriculture, College of Veterinary Medicine, China Agricultural University, 100193, Beijing, People's Republic of China aut NATIONALLICENCE 700 1- Wang Hui Key Laboratory of Animal Epidemiology and Zoonosis, Ministry of Agriculture, College of Veterinary Medicine, China Agricultural University, 100193, Beijing, People's Republic of China aut NATIONALLICENCE 700 1- Sun Zhenhong Taishan Medical University, 271000, Taian, Shandong, People's Republic of China aut NATIONALLICENCE 700 1- He Jinyan Laboratory of Rapid Diagnostic Technology for Animal Disease, College of Veterinary Medicine, China Agricultural University, 100193, Beijing, People's Republic of China aut NATIONALLICENCE 700 1- Shen Junjun Laboratory of Rapid Diagnostic Technology for Animal Disease, College of Veterinary Medicine, China Agricultural University, 100193, Beijing, People's Republic of China aut NATIONALLICENCE 700 1- Wu Wenxue Laboratory of Rapid Diagnostic Technology for Animal Disease, College of Veterinary Medicine, China Agricultural University, 100193, Beijing, People's Republic of China aut NATIONALLICENCE 700 1- Li Jinxiang Chinese Academy of Agricultural Sciences, 100081, Beijing, People's Republic of China aut NATIONALLICENCE 773 0- Applied Microbiology and Biotechnology Springer Berlin Heidelberg 99/4(2015-02-01), 1859-1871 0175-7598 99:4<1859 2015 99 253