Abstract
Begomoviruses interfere with host plant machinery to evade host defense mechanism by interacting with plant proteins. In the old world, this group of viruses are usually associated with betasatellite that induces severe disease symptoms by encoding a protein, βC1, which is a pathogenicity determinant. Here, we show that βC1 encoded by Cotton leaf curl Multan betasatellite (CLCuMB) requires Gossypium hirsutum calmodulin-like protein 11 (Gh-CML11) to infect cotton. First, we used the in silico approach to predict the interaction of CLCuMB-βC1 with Gh-CML11. A number of sequence- and structure-based in-silico interaction prediction techniques suggested a strong putative binding of CLCuMB-βC1 with Gh-CML11 in a Ca+2-dependent manner. In-silico interaction prediction was then confirmed by three different experimental approaches: The Gh-CML11 interaction was confirmed using CLCuMB-βC1 in a yeast two hybrid system and pull down assay. These results were further validated using bimolecular fluorescence complementation system showing the interaction in cytoplasmic veins of Nicotiana benthamiana. Bioinformatics and molecular studies suggested that CLCuMB-βC1 induces the overexpression of Gh-CML11 protein and ultimately provides calcium as a nutrient source for virus movement and transmission. This is the first comprehensive study on the interaction between CLCuMB-βC1 and Gh-CML11 proteins which provided insights into our understating of the role of βC1 in cotton leaf curl disease.
Highlights
Geminiviridae is one of the largest groups of plant viruses
TYLCCNB was shown to interact with CML protein in N. benthamiana, but TYLCCNB-βC1 shares only 30% identity with Cotton leaf curl Multan betasatellite (CLCuMB)-βC1 S2 Table
This sequence to structure analysis data suggested that CLCuMB-βC1 may or may not have the binding ability with Gh-CML11 protein which is in need to be investigated independently for CLCuMV role in the presence of calmodulin like protein
Summary
Geminiviridae is one of the largest groups of plant viruses. Members in this family possess circular, single-stranded (ss) DNA as their genome, and infect economically important crops such as cotton, cassava, grains, maize, and vegetables [1]. CLCuMB-βC1 interaction with Gh-CML11 in CLCuD and interfaces”. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript
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