Spatial layout of chromosomes enables equal allocation of genetic material during cell division. Congression of chromosomes that are positioned on the poles of the mitotic spindle is mediated by kinesin-7/CENP-E, a microtubule plus-end directed motor protein. Currently, there are two models that explain how CENP-E directs chromosomes exclusively towards cell equator. In the first model, CENP-E drives chromosome movements from high Aurora A kinase concentration on the poles, towards the cell equator, then binds protein phosphatase 1 which dephosphorylates it and enables stable binding of kinetochores to microtubules. The second model is based on different preferences of two motor proteins with opposite directionalities, namely CENP-E and dynein, for detyrosinated and tyrosinated microtubules, respectively, which control the directionality of chromosome movements during cell division. However, the relationship of individual contributions of activity regulation of CENP-E by phosphorylation and microtubule detyrosination remains to be elucidated. This set of experiments focused on clarifying the role of CENP-E phosphorylation by Aurora A and B and its impact on CENP-E-dynein activity switch and potential interaction. The hypothesis is that T422 phosphorylation of CENP-E prevents its premature stripping by Dyenin from bioriented chromosomes’ kinetochores, thus facilitating the accurate cell division. The methods used were molecular cloning, lentiviral infection, immunofluorescence following protein inhibition and siRNA knock down, confocal microscopy, Western blot and immunoprecipitation.