Elucidation of functional domains of Chandipura virus Nucleocapsid protein involved in oligomerization and RNA binding: implication in viral genome encapsidation.
Chandipura virus, a member of the vesiculovirus genera, has been recently recognized as an emerging human pathogen. Previously, we have shown that Chandipura virus Nucleocapsid protein N is capable of binding to both specific viral leader RNA as well as non-viral RNA sequences, albeit in distinct monomeric and oligomeric states, respectively. Here, we distinguish the regions of N involved in oligomerization and RNA binding using a panel of deletion mutants. We demonstrate that deletion in the N-terminal arm completely abrogates self-association of N protein. Monomer N specifically recognizes viral leader RNA using its C-terminal 102 residues, while oligomerization generates an additional RNA binding surface involving the N-terminal 320 amino acids of N overlapping with a protease resistant core that is capable of forming nucleocapsid like structure and also binding heterogeneous RNA sequences. Finally, we propose a model to explain the mechanism of genome encapsidation of this important human pathogen.
Mondal
Arindam
A
Department of Biochemistry, University of Calcutta, Kolkata, India.
Bhattacharya
Raja
R
Ganguly
Tridib
T
Mukhopadhyay
Subhradip
S
Basu
Atanu
A
Basak
Soumen
S
Chattopadhyay
Dhrubajyoti
D
D006801:Humans; D046529:Microscopy, Electron, Transmission; D019590:Nucleocapsid Proteins / Q000235:genetics / Q000378:metabolism*; D011485:Protein Binding; D055503:Protein Multimerization; D017434:Protein Structure, Tertiary; D012367:RNA, Viral / Q000378:metabolism*; D016601:RNA-Binding Proteins / Q000235:genetics / Q000378:metabolism*; D017384:Sequence Deletion; D018116:Vesiculovirus / Q000235:genetics / Q000502:physiology*; D014771:Virion / Q000648:ultrastructure; D019065:Virus Assembly
D019590:Nucleocapsid Proteins; D012367:RNA, Viral; D016601:RNA-Binding Proteins