The Establishment Of Large-scale Yeast Two-hybrid Technology And Human The Zak Beta And Ywhaz Protein Interaction

This research successfully establish a standard method for two step-mating yeast two-hybrid system,and a large-scale screen platform for protein-protein interactions for the first time.This platform will make effective influence on the large-scale proteomics research in the future.Then we find out 163 pairs of interactions for the first time between more than 1000 human liver proteins.These data provides a sound basis for future deeper research.MAPK signal transduction pathway is a very important one in eukaryote cells,serine/threonine kinase ZAK located at the upstream of this pathway which may linking to many unclear function mechanism of the MAPK pathway. Our research use the newly established high through-put yeast two hybrid platform to do large scale mating with the library of 1000 yeast clones, containing genes of human liver proteins,by using Human ZAKβas a bait. YWHAZ was identified to interact with ZAKβfor the first time.Pull-down and coimmunoprecipitation were used to confirm this interaction in vitro and in vivo.Moreover,a co-localization assay showed that the two can co-localizes in the cytoplasm,through the western blot using the anti-phosphorylation antibody,we find out that ZAKβcan phosphotylate YWHAZ.At last,we discovered that this interaction can increase the G2 phase level in cell cycle.In addition,the article provides clues for further research of this interaction on its potencial biological function.