Journal de protéomique et bioinformatique
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Molecular Cloning of MAP Kinase Genes and In silico Identification of their Downstream Transcription Factors Involved in Pathogenesis of Karnal bunt(Tilletia indica) of Wheat

Atul K Gupta, Anshita Goel, JM Seneviratne, GK Joshi and Anil Kumar

The mitogen-activated protein kinase (MAPK) pathways control diverse cellular functions in pathogenic fungi, including sexual differentiation, stress response, and maintenance of cell wall integrity. Here we identified and characterized T indica MAPK genes, which are homologues to the other fungi viz. Pmk1 (Pathogenicity MAP Kinase-1) and Fus3 that is known to play an important role in pathogenicity and mating pheromone response during fungal development. It was performed through gene specific genomic DNA amplification, cloning, sequencing, homology match, annotation, motif analysis, structure prediction and protein-protein interaction. Comparative genomics approaches were applied for studying the role of such genes involved in pathogenesis. These analyses give new insights into similarities and dissimilarities as well as specific characteristics of T indica genomes when compared with Saccharomyces cerevisae and Ustilago maydis . Multiple sequence alignment and motif analysis of these genes revealed high conserveness. The result of structure prediction showed that the models are authentic with 88.9% residues are found in most favored regions in TiFus3 while 83.1% residues in TiPmk1. Key functional regulations of these genes involved numerous biological processes including fungal development and pathogenesis can also be achieved through understanding of protein- protein interaction using a downstream target Ste12p transcription factor. Interaction between two different pairs of protein i.e. TiPmk1 with Ste12p and TiFus3 with Ste12p showed that TiPmk1 interact with Ste12p with -551.19 kcal total energy while TiFus3 interact with Ste12p -157.76 kcal total energy. There is currently no direct evidence for any direct activation of this transcription factor by MAP kinase in T indica , in silico interaction study provides ample opportunities to validate its role in the pathogenesis and fungal development. Thus, it will ultimately lead to help in the development of novel strategies for controlling the Karnal bunt (KB) disease of wheat, incited by T indica.