Kinetics of interaction of Cotton Leaf Curl Kokhran Virus-Dabawali (CLCuKV-Dab) coat protein and its mutants with ssDNA

Priyadarshini, Poornima CG and Savithri, HS (2009) Kinetics of interaction of Cotton Leaf Curl Kokhran Virus-Dabawali (CLCuKV-Dab) coat protein and its mutants with ssDNA. In: Virology, 386 (2). pp. 427-437.

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Abstract

Gemini viral assembly and transport of viral DNA into nucleus for replication, ssentially involve DNA-coat protein interactions. The kinetics of interaction of Cotton LeafCtirl Kokhran Virus-Dabawali recombinant coat protein (rCP) with DNA was studied by electrophoretic mobility shift assay (EMSA) and Surface plasmon resonance (SPR). The rCP interacted with ssDNA with a K-A, of 2.6 +/- 0.29 x 10(8) M-1 in a sequence non-specific manner. The CP has a conserved C2H2 type zinc finger motif composed of residues C68, C72, H81 and H85. Mutation of these residues to alanine resulted in reduced binding to DNA probes. The H85A mutant rCP showed the least binding with approximately 756 fold loss in the association rate and a three order magnitude decrease in the binding affinity as compared to rCP. The CP-DNA interactions via the zinc finger motif could play a Crucial role ill Virus assembly and in nuclear transport. (C) 2009 Elsevier Inc.

Item Type:	Journal Article
Publication:	Virology
Publisher:	Elsevier science
Additional Information:	Copyright for this article belongs to Elsevier science.
Keywords:	Geminiviridae;Begomoviruses;Cotton Leaf Curl Kokhran Virus (CLCuKV);Coat protein;Surface plasmon resonance;ssDNA interaction;Zinc finger. motif
Department/Centre:	Division of Biological Sciences > Biochemistry
Date Deposited:	27 Aug 2009 06:36
Last Modified:	19 Sep 2010 05:37
URI:	http://eprints.iisc.ac.in/id/eprint/21461

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