Ebola virus nucleoprotein (eNP) assembles into higher-ordered structures that form the viral nucleocapsid (NC) and serve as the scaffold for viral RNA synthesis. However， molecular insights into the NC?assembly process are lacking. Using a hybrid approach， we characterized the NC-like assembly of eNP， identified novel regulatory elements， and described how these elements impact function. We generated a three-dimensional structure of the eNP NC-like assembly at 5.8?? using electron cryo-microscopy and identified a new regulatory role for?eNP helices α22-α23. Biochemical， biophysical， and mutational analyses revealed that inter-eNP contacts within α22-α23 are critical for viral NC assembly and reg... More
Ebola virus nucleoprotein (eNP) assembles into higher-ordered structures that form the viral nucleocapsid (NC) and serve as the scaffold for viral RNA synthesis. However， molecular insights into the NC?assembly process are lacking. Using a hybrid approach， we characterized the NC-like assembly of eNP， identified novel regulatory elements， and described how these elements impact function. We generated a three-dimensional structure of the eNP NC-like assembly at 5.8?? using electron cryo-microscopy and identified a new regulatory role for?eNP helices α22-α23. Biochemical， biophysical， and mutational analyses revealed that inter-eNP contacts within α22-α23 are critical for viral NC assembly and regulate viral RNA synthesis. These observations suggest that the N terminus and α22-α23 of eNP function as context-dependent regulatory modules (CDRMs). Our current study provides a framework for a structural mechanism for NC-like assembly and a new therapeutic target.
