Ebola virus （EBOV） harbors an RNA genome encapsi- dated by nucleoprotein （NP） along with other viral pro- teins to form a nucleocapsid complex. Previous Cryo- eletron tomography and biochemical studies have shown ...Ebola virus （EBOV） harbors an RNA genome encapsi- dated by nucleoprotein （NP） along with other viral pro- teins to form a nucleocapsid complex. Previous Cryo- eletron tomography and biochemical studies have shown the helical structure of EBOV nucleocapsid at nanometer resolution and the first 450 amino-acid of NP （NP△451-739） alone is capable of forming a helical nucleocapsid-Iike complex （NLC）. However, the struc- tural basis for NP-NP interaction and the dynamic pro- cedure of the nucleocapsid assembly is yet poorly understood. In this work, we, by using an E. coli expression system, captured a series of images of NP△451-739 conformers at different stages of NLC assembly by negative-stain electron microscopy, which allowed us to picture the dynamic procedure of EBOV nucleocapsid assembly. Along with further biochemical studies, we showed the assembly of NLC is salt-sensi- tive, and also established an indispensible role of RNA in this process. We propose the diverse modes of NLC elongation might be the key determinants shaping the plasticity of EBOV virions. Our findings provide a new model for characterizing the self-oligomerization of viral nucleoproteins and studying the dynamic assembly process of viral nucleocapsid in vitro.展开更多
基金We would like to thank Jingnan Liang for technical assistance in electron microscopy, the core facility of Institute of Microbiology, Chinese Academy of Sciences. This work was supported by the National Natural Science Foundation of China （Grant No. 81590761）, the National Basic Research Program （973 Program） （Nos. 2013CB531502 and 2014CB542503）, and Strategic Priority Research Program of the Chinese Academy of Sciences （XDB08020100）.
文摘Ebola virus （EBOV） harbors an RNA genome encapsi- dated by nucleoprotein （NP） along with other viral pro- teins to form a nucleocapsid complex. Previous Cryo- eletron tomography and biochemical studies have shown the helical structure of EBOV nucleocapsid at nanometer resolution and the first 450 amino-acid of NP （NP△451-739） alone is capable of forming a helical nucleocapsid-Iike complex （NLC）. However, the struc- tural basis for NP-NP interaction and the dynamic pro- cedure of the nucleocapsid assembly is yet poorly understood. In this work, we, by using an E. coli expression system, captured a series of images of NP△451-739 conformers at different stages of NLC assembly by negative-stain electron microscopy, which allowed us to picture the dynamic procedure of EBOV nucleocapsid assembly. Along with further biochemical studies, we showed the assembly of NLC is salt-sensi- tive, and also established an indispensible role of RNA in this process. We propose the diverse modes of NLC elongation might be the key determinants shaping the plasticity of EBOV virions. Our findings provide a new model for characterizing the self-oligomerization of viral nucleoproteins and studying the dynamic assembly process of viral nucleocapsid in vitro.
