Journal Article

Tomato leaf curl Kerala virus (ToLCKeV) AC3 protein forms a higher order oligomer and enhances ATPase activity of replication initiator protein (Rep/AC1)

Mukherjee, Sunil K, Choudhury, Nirupam R, Pasumarthy, Kalyan K

2010

Overview

Background Geminiviruses are emerging plant viruses that infect a wide variety of vegetable crops, ornamental plants and cereal crops. They undergo recombination during co-infections by different species of geminiviruses and give rise to more virulent species. Antiviral strategies targeting a broad range of viruses necessitate a detailed understanding of the basic biology of the viruses. ToLCKeV, a virus prevalent in the tomato crop of Kerala state of India and a member of genus Begomovirus has been used as a model system in this study. Results AC3 is a geminiviral protein conserved across all the begomoviral species and is postulated to enhance viral DNA replication. In this work we have successfully expressed and purified the AC3 fusion proteins from E. coli . We demonstrated the higher order oligomerization of AC3 using sucrose gradient ultra-centrifugation and gel-filtration experiments. In addition we also established that ToLCKeV AC3 protein interacted with cognate AC1 protein and enhanced the AC1-mediated ATPase activity in vitro. Conclusions Highly hydrophobic viral protein AC3 can be purified as a fusion protein with either MBP or GST. The purification method of AC3 protein improves scope for the biochemical characterization of the viral protein. The enhancement of AC1-mediated ATPase activity might lead to increased viral DNA replication.

• Publisher: BioMed Central,Springer Nature B.V,BMC
• Subject: Adenosine Triphosphatases - genetics / Adenosine Triphosphatases - metabolism / Adenosinetriphosphatase / Begomovirus / Begomovirus - chemistry / Begomovirus - enzymology / Begomovirus - genetics / Begomovirus - metabolism / Biomedical and Life Sciences / Biomedicine / Cereals / Crops / DNA biosynthesis / DNA Helicases - genetics / DNA Helicases - metabolism / DNA replication / E coli / Escherichia coli / Experiments / Fusion protein / gel chromatography / genus / Hydrophobicity / India / Infections / leaf curling / Leaves / Lycopersicon esculentum / Microbiology / Molecular weight / Oligomerization / Ornamental plants / Plant viruses / Protein Binding / Protein Conformation / protein purification / Proteins / purification methods / Recombination / Regression analysis / Replication / replication initiator protein / species / Studies / Sucrose / tomatoes / Trans-Activators - genetics / Trans-Activators - metabolism / Vegetables / Viral infections / Viral Proteins - chemistry / Viral Proteins - genetics / Viral Proteins - metabolism / Virology / virulence / viruses