CENP-A phosphorylation by Aurora-A in prophase is required for enrichment of Aurora-B at inner centromeres and for kinetochore function.
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.
Kunitoku
Naoko
N
Department of Tumor Genetics and Biology, Graduate School of Medical Sciences, Kumamoto University, 1-1-1 Honjo, Kumamoto 860-8556, Japan.
Sasayama
Takashi
T
Marumoto
Tomotoshi
T
Zhang
Dongwei
D
Honda
Shinobu
S
Kobayashi
Osamu
O
Hatakeyama
Katsuyoshi
K
Ushio
Yukitaka
Y
Saya
Hideyuki
H
Hirota
Toru
T
D000595:Amino Acid Sequence; D064107:Aurora Kinase B; D064127:Aurora Kinases; D001324:Autoantigens; D002467:Cell Nucleus / Q000378:metabolism; D000076247:Centromere Protein A; D002868:Chromosomal Proteins, Non-Histone / Q000235:genetics / Q000378:metabolism*; D006367:HeLa Cells; D006801:Humans; D047908:Intracellular Signaling Peptides and Proteins; D007668:Kidney / Q000166:cytology; D018386:Kinetochores / Q000201:enzymology*; D008677:Metaphase / Q000502:physiology; D008870:Microtubules / Q000502:physiology; D008938:Mitosis / Q000502:physiology; D008969:Molecular Sequence Data; D009687:Nuclear Proteins / Q000235:genetics / Q000378:metabolism*; D010766:Phosphorylation; D011418:Prophase / Q000502:physiology; D017346:Protein Serine-Threonine Kinases / Q000378:metabolism*; D011506:Proteins; D034741:RNA, Small Interfering; D012694:Serine / Q000378:metabolism
D001324:Autoantigens; C000619700:CENPA protein, human; D000076247:Centromere Protein A; D002868:Chromosomal Proteins, Non-Histone; D047908:Intracellular Signaling Peptides and Proteins; D009687:Nuclear Proteins; D011506:Proteins; D034741:RNA, Small Interfering; C103265:aryl hydrocarbon receptor-interacting protein; D012694:Serine; C576921:AURKB protein, human; D064107:Aurora Kinase B; D064127:Aurora Kinases; D017346:Protein Serine-Threonine Kinases