RESUMO
This study demonstrates for the first time that the matrix (M) protein of BEFV is a nuclear targeting protein that shuttles between the nucleus and the cytoplasm in a transcription-, carrier-, and energy-dependent manner. Experiments performed in both intact cells and digitonin-permeabilized cells revealed that M protein targets the nucleolus and requires carrier, cytosolic factors or energy input. By employing sequence and mutagenesis analyses, we have determined both nuclear localization signal (NLS) 6KKGKSK11 and nuclear export signal (NES) 98LIITSYL TI106 of M protein that are important for the nucleocytoplasmic shuttling of M protein. Furthermore, we found that both lamin A/C and chromosome maintenance region 1 (CRM-1) proteins could be coimmunoprecipitated and colocalized with the BEFV M protein. Knockdown of lamin A/C by shRNA and inhibition of CRM-1 by leptomycin B significantly reduced virus yield. Collectively, this study provides novel insights into nucleocytoplasmic shuttling of the BEFV M protein modulated by lamin A/C and CRM-1 and by a transcription- and carrier- and energy-dependent pathway.
Assuntos
Transporte Ativo do Núcleo Celular , Vírus da Febre Efêmera Bovina , Lamina Tipo A , Sinais de Localização Nuclear , Animais , Transporte Ativo do Núcleo Celular/genética , Núcleo Celular/metabolismo , Cromossomos/metabolismo , Citoplasma/metabolismo , Lamina Tipo A/genética , Lamina Tipo A/metabolismo , Vírus da Febre Efêmera Bovina/metabolismo , Proteínas Estruturais Virais/metabolismoRESUMO
The purpose of this study was to investigate the antibody response to the bovine ephemeral fever virus (BEFV) vaccine in Japanese Black calves. Twenty-eight Japanese Black calves, which were raised on an ordinal farm, were divided into two groups. Fifteen calves received the inactivated BEFV vaccine at 12 and 16 weeks of age (vaccination group), and 13 calves did not receive the vaccine (non-vaccination group). Blood samples were obtained at 0, 4, 8, 12, 16, 20, 24, 28, and 32 weeks of age. As the results, in the vaccination group, the antibody titers at 16, 20, 24, 28, and 32 weeks of age were significantly higher than those at 0, 4, 8, and 12 weeks of age (p < 0.01). Additionally, antibody titer in the vaccination group increased after 16 weeks of age and showed a significantly higher level than that in the non-vaccination group throughout the remaining experimental period (p < 0.01). These results might be helpful in establishing a vaccination program against BEFV in calves.