An intermediate region in C-terminal of phosphoprotein is required for binding to nucleocapsid of Newcastle disease virus
The phosphoprotein (P) of paramyxoviruses plays a central role in the viral genome replication and transcription. The P either binds to the unassembled nucleocapsid (NP0) to assist in viral genomic replication or binds to assembled NP (NP-RNA or NPNC) to transcribe genome to produce the subgenomic...
| Main Authors: | , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Academic Journals
2011
|
| Subjects: | |
| Online Access: | http://irep.iium.edu.my/18451/ http://irep.iium.edu.my/18451/ http://irep.iium.edu.my/18451/1/An_intermediate.pdf |
| Summary: | The phosphoprotein (P) of paramyxoviruses plays a central role in the viral genome replication and
transcription. The P either binds to the unassembled nucleocapsid (NP0) to assist in viral genomic
replication or binds to assembled NP (NP-RNA or NPNC) to transcribe genome to produce the subgenomic
mRNAs. In this study, the region of P that binds to NPNC was mapped. To determine the
binding region, 18 N- and C-terminally truncated P mutants were synthesized by in vitro translation in
rabbit reticulocytes and mixed with purified NP (NPNC). The mutants which did not bind to NP were
considered as mutants and they contain deleted regions that may be involved in binding to NP. To
identify the mutants that did not bind to NP, radioimmunoprecipitation and protein binding assays were
used. Based on radioimmunoprecipitation analysis, it was shown that the region of P that binds to NP is
located within the internal region of C-terminal half of P, from amino acids 243 to 279. In agreement with
the radioimmunoprecipitation analysis, protein binding assay showed that the interactive domain was
mapped to the internal region of the C-terminal half of P. However, a slightly bigger region of interactive
domain (amino acids 224 to 279) was determined by the latter assay. As protein binding assay is
considered as a more sensitive assay as compared to radioimmunoprecipitation assay, thus, the
interactive region of P to NPNC was located within C-terminal half of P between amino acids 224 to 279. |
|---|