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Yellow rust, caused by Puccinia striiformis Westend. f. sp. tritici Eriksson is one of the most severe leaf diseases of wheat. Aim of this study is to illuminate the downstream signaling pathways upon incompetible infection of rust pathogen in wheat, thus to understand the genes involved in resistance mechanism. The strategy used is the dual bait yeast two-hybrid analysis which is the most powerful method for in vivo detection of protein-protein interactions. The bait proteins used are / the domains of Yr10 yellow rust resistance gene, Rad6 gene which is considered to have a critical role in R gene mediated signaling pathway, and WR5 gene fragment which is an unknown protein having homology to the WD40 repeat containing protein with apoptosis related activity. Screening of a yeast prey library with these baits revealed proteins having mostly apoptosis related functions (SRP72, POR1, CSE1), translation initiation control in response to stress conditions (Gcn2p, Eap1p), phosphorylation (SKY1) and dephosphorylation activities (GAC1), cell cycle control (FAR1), oxidative stress control (OXR1), protein degradation control (TOM1), protein folding control (CPR7) and ion homeostasis in the cell (POR1, GAC1). The significance of the study can be summarized as i) being the first yeast two hybrid analysis of a wheat R gene, ii) being able to detect interacting partners with anticipated functions, iii) most importantly, initiating further detailed analysis of the key interactors.