Abstract
Cotton leaf curl disease (CLCuD) caused by viruses of genus Begomovirus is a major constraint to cotton (Gossypium hirsutum) production in many cotton-growing regions of the world. Symptoms of the disease are caused by Cotton leaf curl Multan betasatellite (CLCuMB) that encodes a pathogenicity determinant protein, βC1. Here, we report the identification of interacting regions in βC1 protein by using computational approaches including sequence recognition, and binding site and interface prediction methods. We show the domain-level interactions based on the structural analysis of G. hirsutum SnRK1 protein and its domains with CLCuMB-βC1. To verify and validate the in silico predictions, three different experimental approaches, yeast two hybrid, bimolecular fluorescence complementation and pull down assay were used. Our results showed that ubiquitin-associated domain (UBA) and autoinhibitory sequence (AIS) domains of G. hirsutum-encoded SnRK1 are involved in CLCuMB-βC1 interaction. This is the first comprehensive investigation that combined in silico interaction prediction followed by experimental validation of interaction between CLCuMB-βC1 and a host protein. We demonstrated that data from computational biology could provide binding site information between CLCuD-associated viruses/satellites and new hosts that lack known binding site information for protein–protein interaction studies. Implications of these findings are discussed.
Highlights
Plant viruses cause considerable damage to quality and crop yield and threaten food security in several parts of the world (Oerke and Dehne, 2004)
G. hirsutum-coded SnRK1 protein structure and Cotton leaf curl Multan betasatellite (CLCuMB)-βC1 were used for domain-based interaction prediction, whereas Solanum lycopersicum-coded SnRK1 and Tomato yellow leaf curl China betasatellite (TYLCCNB)-βC1 complex were used as a control
Through multiple lines of evidence, that CLCuMB-βC1 serves as a pathogenicity determinant by interacting with G. hirsutumencoded SnRK1α (GhSnRK1) through ubiquitin-associated domain (UBA) and autoinhibitory sequence (AIS) domains
Summary
Plant viruses cause considerable damage to quality and crop yield and threaten food security in several parts of the world (Oerke and Dehne, 2004). Geminiviruses interact with several proteins in the host to cause changes in their transcription and translation machinery for virus multiplication. Geminivirdae is divided into nine genera based on their genome organization, insect vectors and host range (Martin et al, 2011). Begomovirus is the largest and most economically important genus, and viruses in this genus cause serious diseases in agronomic and horticultural crops such as cotton, cassava, maize, and tomato (Brown et al, 2015). Viruses causing cotton leaf curl disease (CLCuD) are betasatellite-requiring monopartite begomoviruses that cause serious economic damage to cotton (Gossypium hirsutum L.) in the Indian subcontinent and Africa (Nawaz-ulRehman et al, 2009; Tiendrébéogo et al, 2010)
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