PRO11 interacts physically with SmKIN3 (A) and SmKIN24 (B) in a yeast two-hybrid analysis.

<p>Shown are serial dilutions of diploid yeast strains obtained after mating and spread on SD medium lacking tryptophan (trp) and leucine (leu) or trp, leu and adenine (ade) to verify the interaction of both proteins. The left picture displays a control, which ensures presence of both plasmids in the diploid strain; trp and leu prototrophy are regained by genes present on the plasmid. The right picture displays the interaction assay; ade prototrophy is obtained by positive interaction between the proteins fused to GAL4 binding domain (BD) and GAL4 activation domain (AD). Full length versions of SmKIN3 and SmKIN24 without any introns were tested. GAL4-binding domain was fused to SmKIN3 or SmKIN24 GAL4 activation domain to PRO11. Reverse application of activation and binding domain was not possible due to transactivation of BD-PRO11. Yeast transformants expressing genes coding for BD-SmKIN3 or BD-SmKIN24 and AD-RanBPM served as positive control (46). As negative control a diploid strain carrying empty vectors pGADT7 (AD) and pGBKT7 (BD) was used. No interaction between SmMOB3 and SmKIN3 or SmKIN24 has been observed.