Significance In mitosis, microtubules extend and shrink before the bilateral attachment is established. However, which molecules regulate this activity for spindle formation is not fully elucidated. Using two in-house developed small molecules that target the Aurora kinases, we show that hepatoma up-regulated protein (HURP) is highly dynamic, trafficking between centrosome and kinetochore driven by Aurora A-dependent phosphorylation and protein phosphatase 1/2A-associated dephosphorylation. These compounds demonstrate a spatial hierarchical preference of HURP in the attachment of microtubules extending from the mother to the daughter centrosome. These findings help explain the biology of mitosis and may lead to the development of anticancer compounds.