Domains of Rinderpest Virus Phosphoprotein Involved in Interaction with Itself and the Nucleocapsid Protein

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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Abstract

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
Publication:	Virology
Publisher:	Academic Press
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
URI:	http://eprints.iisc.ac.in/id/eprint/12086

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