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Indian Journal of Virology
Year : 2008, Volume : 19, Issue : 1

Print ISSN : 0970-2822.

P-68. Functional analysis of coat protein and AV2 protein in the movement of Cotton Leaf Curl Kokhran virus-Dabawali (CLCuKV-Dab), a monopartite begomo-virus

Priyadarshini Poornima C.G., Thippeswamy R., Savithri H.S.

Department of Biochemistry, Indian Institute of Science, Bangalore-560012, India.

Abstracts of the papers presented at the International Conference of Indian Virological Society on “Emerging and Re-emerging viral Diseases of the Tropics and Subtropics” at Indian Agricultural Research Institute, New Delhi, India, December 11–14, 2007.

Summary

Plant viruses must cross cell wall barrier posed by the plants in order move and cause the systemic infection. These processes are governed both by viral proteins and host factors which exploit many complex mechanisms like opening of plasmodesmata and formation of plasma membrane-lined channels. In contrast to most RNA viruses, DNA viruses replicate in the nucleus and hence employ additional mechanisms to facilitate their movement. Begomoviruses, a subgenera in the family of Geminiviridae encode two non-structural movement proteins NSP and MP to cross nuclear envelope and the plasma membrane. Monopartite begomoviruses lack DNA-B and hence the NSP and MP proteins. It has been suggested that the CP and AV2 proteins encoded by DNA-A could replace the functions of NSP and MP respectively. In the present study, coat protein (CP) encoded by AV1 ORF and the putative viral movement protein encoded by AV2 ORF of Cotton Leaf curl Kokhran virus-Dabawali (CLCuKV-Dab) were functionally characterized. The two proteins were cloned and expressed in insect cells using the baculovirus expression system as C-terminal fusion of green fluorescent proteins. The cells after the infection were used for in vivo localization experiments by confocal microscopy. The CP localized in the nucleus where as AV2 localized in the cell periphery. Subcellular fractionation and subsequent western blot analysis confirmed these observations. Also, in vitro protein-protein interactions and DNA binding studies were performed with bacterially expressed CP and AV2. These results confirm that CP and AV2 proteins are involved in cell to cell transport of viral DNA.

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