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 |
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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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