Original Research ARTICLE
In Silico Prediction and Validations of Domains Involved in Gossypium hirsutum SnRK1 Protein Interaction with Cotton leaf curl Multan betasatellite encoded βC1
- 1National Institute for Biotechnology and Genetic Engineering, Pakistan
- 2Pakistan Institute of Engineering and Applied Sciences, Pakistan
- 3Department of Biology, University of Washington, United States
- 4Washington State University, United States
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 first 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. These 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.
Keywords: Cotton leaf curl disease, sucrose-nonfermenting 1 (SNF1) kinase, yeast two hybrid, bimolecular fluorescence complementation, Pull down assay, Begomovirus, Geminivirus, Cotton leaf curl Multan betasatellite
Received: 27 Dec 2018;
Accepted: 01 May 2019.
Edited by:Jeremy Astier, INRA UMR1347 Agroécologie, France
Reviewed by:Jin Koh, University of Florida, United States
C N L. Reddy, University of Agricultural Sciences, Bangalore, India
Copyright: © 2019 Kamal, Minhas, Farooq, Tripathi, Hamza, Mustafa, Khan, Mansoor, Pappu and Amin. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
* Correspondence: Dr. Imran Amin, National Institute for Biotechnology and Genetic Engineering, Faisalabad, Pakistan, email@example.com