Ensemble Structure of the Modular and Flexible Full-Length Vesicular Stomatitis Virus Phosphoprotein

The phosphoprotein (P) is an essential component of the viral replication machinery of non-segmented negative‐strand RNA viruses, connecting the viral polymerase to its nucleoprotein–RNA template and acting as a chaperone of the nucleoprotein by preventing nonspecific encapsidation of cellular RNAs. The phosphoprotein of vesicular stomatitis virus (VSV) forms homodimers and possesses a modular organization comprising two stable, well-structured domains concatenated with two intrinsically disordered regions. Here, we used a combination of nuclear magnetic resonance spectroscopy and small-angle X-ray scattering to depict VSV P as an ensemble of continuously exchanging conformers that captures the dynamic character of this protein. We discuss the implications of the dynamics and the large conformational space sampled by VSV P in the assembly and functioning of the viral transcription/replication machinery. [Display omitted] ► Combined nuclear magnetic resonance and small‐angle X‐ray scattering approaches to characterize the structure of VSV phosphoprotein. ► The structure is described as a conformational ensemble. ► Roles are proposed for the flexibility of VSV phosphoprotein. ► New insights about the structural organization of the replication complex are gained.