Shaji, Daniel and Shaila, MS (1999) Domains of Rinderpest Virus Phosphoprotein Involved in Interaction with Itself and the Nucleocapsid Protein. In: Virology, 258 (2). pp. 415-424.
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The yeast two-hybrid system was used to identify domains involved in specific in vivo interactions between the Rinderpest virus (RPV) phosphoprotein (P) and nucleocapsid protein (N). N and P genes were cloned in both the yeast GAL4 DNA-binding and GAL4 activation domain vectors, which enabled analysis of self and interprotein interactions. Mapping of the domain of P protein involved in its association with itself revealed that the COOH-terminal 32 amino acids (316–347) that forms a part of the highly conserved coiled coil region is important for interaction. In addition, just the coiled coil region of RPV P protein fused to the DNA-binding domain and activation domain of GAL4 was found to be sufficient to bring about activation of the $\beta-$galactosidase reporter. Similarly, mapping of the domains of P protein involved in its interaction with N protein revealed that $NH_2-$terminal 59 amino acids and COOH-terminal 32 amino acids (316–347) involved in P–P interaction are simultaneously required for association with N protein. Interestingly, a P protein mutant with just the $NH_2-$terminal 59 amino acids and the coiled coil domain with all other P protein regions deleted retained its ability to interact with N protein. Furthermore, we were able to show N and P protein interaction in vitro using recombinant N and P proteins expressed in Escherichia coli, demonstrating the existence of direct physical interaction between the two proteins.
|Item Type:||Journal Article|
|Additional Information:||Copyright of this article belongs to Academic Press.|
|Keywords:||Rinderpest virus;Phosphoprotein;Nucleocapsid protein;Yeast two hybrid system;Interaction|
|Department/Centre:||Division of Biological Sciences > Microbiology & Cell Biology|
|Date Deposited:||05 Oct 2007|
|Last Modified:||19 Sep 2010 04:40|
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