Nucleocapsid (NP) and Phospho-(P) Proteins of Newcastle Disease Virus: Identification of Regions on NP That Form Particles and Interact with P
The nucleocapsid protein ( NP) of Newcastle disease virus (NDV) plays an important role in the replication of the viral genomic RNA. The NP is closely associated with the viral phosphoprotein ( P) and this association is crucial in ensuring the specific binding ofNP to the viral RNA. In order to...
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| Format: | Thesis |
| Language: | English English |
| Published: |
2003
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| Subjects: | |
| Online Access: | http://psasir.upm.edu.my/id/eprint/9561/1/FSAS_2003_21_A.pdf |
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| Summary: | The nucleocapsid protein ( NP) of Newcastle disease virus (NDV) plays an
important role in the replication of the viral genomic RNA. The NP is closely
associated with the viral phosphoprotein ( P) and this association is crucial in
ensuring the specific binding ofNP to the viral RNA. In order to have a better
understanding of the structure and functions of the NP, deletion mutagenesis
was carried out to characterise and localise regions involved in NP-NP and
NP-P interactions.
The NP and a fusion derivative (NPcfus) containing a hexa histidine tag
at its C-terminus were produced abundantly in Escherichia coli. These proteins
were fractionated on sucrose gradient centrifugation and microscopic analysis
showed that both the NP and NP cfus proteins self-assembled predominantly into
ring-like particles with the diameter of 24 ± 2 nm around a central hole of 7 ±
1 nm. Some of these ring-like particles stacked together to form herringbonelike
particles which are heterogenous in length with a diameter of 20 ± 2 nm and a central hollow of 5 ± 1 nm. Fusion of the C-terminal end to 29 amino
acids inclusive of the myc epitope and His-tag did not impair ring assembly but
inhibit the formation of the long herringbone particles. Immunogold labelling
of the ring-like particles with the anti-myc antibody showed that the Cterminus
of the NPcfus protein is exposed on the surface of the particles. |
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