CENP-A Phosphorylation by Aurora-A in Prophase Is Required for Enrichment of Aurora-B at Inner Centromeres and for Kinetochore Function

• Published in: Developmental cell Vol. 5; no. 6; pp. 853 - 864
• Main Authors: Kunitoku, Naoko, Sasayama, Takashi, Marumoto, Tomotoshi, Zhang, Dongwei, Honda, Shinobu, Kobayashi, Osamu, Hatakeyama, Katsuyoshi, Ushio, Yukitaka, Saya, Hideyuki, Hirota, Toru
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 01.12.2003
• Subjects: Amino Acid Sequence; Aurora Kinase B; Aurora Kinases; Aurora-A protein; Aurora-B protein; Autoantigens; Cell Nucleus - metabolism; CENP-A protein; Centromere Protein A; Chromosomal Proteins, Non-Histone - genetics; Chromosomal Proteins, Non-Histone - metabolism; HeLa Cells; Humans; Intracellular Signaling Peptides and Proteins; Kidney - cytology; Kinetochores; Kinetochores - enzymology; Metaphase - physiology; Microtubules - physiology; Mitosis - physiology; Molecular Sequence Data; Nuclear Proteins - genetics; Nuclear Proteins - metabolism; Phosphorylation; Prophase - physiology; Protein-Serine-Threonine Kinases - metabolism; Proteins; RNA, Small Interfering; Serine - metabolism
• ISSN: 1534-5807; 1878-1551
• DOI: 10.1016/S1534-5807(03)00364-2

The Aurora (Ipl1)-related kinases are universal regulators of mitosis. We now show that Aurora-A, in addition to Aurora-B, regulates kinetochore function in human cells. A two-hybrid screen identified the kinetochore component CENP-A as a protein that interacts with Aurora-A. Aurora-A phosphorylated CENP-A in vitro on Ser-7, a residue also known to be targeted by Aurora-B. Depletion of Aurora-A or Aurora-B by RNA interference revealed that CENP-A is initially phosphorylated in prophase in a manner dependent on Aurora-A, and that this reaction appears to be required for the subsequent Aurora-B-dependent phosphorylation of CENP-A as well as for the restriction of Aurora-B to the inner centromere in prometaphase. Prevention of CENP-A phosphorylation also led to chromosome misalignment during mitosis as a result of a defect in kinetochore attachment to microtubules. Our observations suggest that phosphorylation of CENP-A on Ser-7 by Aurora-A in prophase is essential for kinetochore function.