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...

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Main Authors: Ahmad Raus, Raha, Siang, Tan Wen, Ali, Abdul Manaf, Yusoff, Khatijah
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
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spelling iium-184512013-06-27T06:24:33Z http://irep.iium.edu.my/18451/ An intermediate region in C-terminal of phosphoprotein is required for binding to nucleocapsid of Newcastle disease virus Ahmad Raus, Raha Siang, Tan Wen Ali, Abdul Manaf Yusoff, Khatijah TA164 Bioengineering 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. Academic Journals 2011-11-07 Article PeerReviewed application/pdf en http://irep.iium.edu.my/18451/1/An_intermediate.pdf Ahmad Raus, Raha and Siang, Tan Wen and Ali, Abdul Manaf and Yusoff, Khatijah (2011) An intermediate region in C-terminal of phosphoprotein is required for binding to nucleocapsid of Newcastle disease virus. African Journal of Biotechnology, 10 (81). pp. 18882-18891. ISSN 1684–5315 http://www.academicjournals.org/AJB/PDF/pdf2011/16DecConf/Raus%20et%20al.pdf
repository_type Digital Repository
institution_category Local University
institution International Islamic University Malaysia
building IIUM Repository
collection Online Access
language English
topic TA164 Bioengineering
spellingShingle TA164 Bioengineering
Ahmad Raus, Raha
Siang, Tan Wen
Ali, Abdul Manaf
Yusoff, Khatijah
An intermediate region in C-terminal of phosphoprotein is required for binding to nucleocapsid of Newcastle disease virus
description 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.
format Article
author Ahmad Raus, Raha
Siang, Tan Wen
Ali, Abdul Manaf
Yusoff, Khatijah
author_facet Ahmad Raus, Raha
Siang, Tan Wen
Ali, Abdul Manaf
Yusoff, Khatijah
author_sort Ahmad Raus, Raha
title An intermediate region in C-terminal of phosphoprotein is required for binding to nucleocapsid of Newcastle disease virus
title_short An intermediate region in C-terminal of phosphoprotein is required for binding to nucleocapsid of Newcastle disease virus
title_full An intermediate region in C-terminal of phosphoprotein is required for binding to nucleocapsid of Newcastle disease virus
title_fullStr An intermediate region in C-terminal of phosphoprotein is required for binding to nucleocapsid of Newcastle disease virus
title_full_unstemmed An intermediate region in C-terminal of phosphoprotein is required for binding to nucleocapsid of Newcastle disease virus
title_sort intermediate region in c-terminal of phosphoprotein is required for binding to nucleocapsid of newcastle disease virus
publisher Academic Journals
publishDate 2011
url http://irep.iium.edu.my/18451/
http://irep.iium.edu.my/18451/
http://irep.iium.edu.my/18451/1/An_intermediate.pdf
first_indexed 2023-09-18T20:27:33Z
last_indexed 2023-09-18T20:27:33Z
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