Effect of osmolytes and chaperone-like action of p-protein on folding of nucleocapsid protein of Chandipura virus

Majumder, Amitabha ; Basak, Soumen ; Raha, Tamal ; Chowdhury, Santanu Pal ; Chattopadhyay, Dhrubajyoti ; Roy, Siddhartha (2001) Effect of osmolytes and chaperone-like action of p-protein on folding of nucleocapsid protein of Chandipura virus Journal of Biological Chemistry, 276 (33). pp. 30948-30955. ISSN 0021-9258

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Official URL: http://www.jbc.org/content/276/33/30948.full

Related URL: http://dx.doi.org/10.1074/jbc.M011705200

Abstract

Amino acid sequences of nucleocapsid proteins are mostly conserved among different rhabdoviruses. The protein plays a common functional role in different RNA viruses by enwrapping the viral genomic RNA in an RNase-resistant form. Upon expression of the nucleocapsid protein alone in COS cells and in bacteria, it forms large insoluble aggregates. In this work, we have reported for the first time the full-length cloning of the N gene of Chandipura virus and its expression in Escherichia coli in a soluble monomeric form and purification using nonionic detergents. The biological activity of the soluble recombinant protein has been tested, and it was found to possess efficient RNA-binding ability. The state of aggregation of the recombinant protein was monitored using light scattering. In the absence of nonionic detergents, it formed large aggregates. Aggregation was significantly reduced in the presence of osmolytes such asd-sorbitol. Aggregate formation was suppressed in the presence of another viral product, phosphoprotein P, in a chaperone-like manner. Both the osmolyte and phosphoprotein P also suppressed aggregation to a great extent during refolding from a guanidine hydrochloride-denatured form. The function of the phosphoprotein and osmolyte appears to be synergistic to keep the N-protein in a soluble biologically competent form in virus-infected cells.

Item Type:Article
Source:Copyright of this article belongs to American Society for Biochemistry and Molecular Biology.
ID Code:10282
Deposited On:04 Nov 2010 06:13
Last Modified:16 May 2016 19:56

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