Mitotic regulation of the stability of microtubule plus-end tracking protein EB3 by ubiquitin ligase SIAH-1 and Aurora mitotic kinases.
Microtubule plus-end tracking proteins (+TIPs) control microtubule dynamics in fundamental processes such as cell cycle, intracellular transport, and cell motility, but how +TIPs are regulated during mitosis remains largely unclear. Here we show that the endogenous end-binding protein family EB3 is stable during mitosis, facilitates cell cycle progression at prometaphase, and then is down-regulated during the transition to G(1) phase. The ubiquitin-protein isopeptide ligase SIAH-1 facilitates EB3 polyubiquitination and subsequent proteasome-mediated degradation, whereas SIAH-1 knockdown increases EB3 stability and steady-state levels. Two mitotic kinases, Aurora-A and Aurora-B, phosphorylate endogenous EB3 at Ser-176, and the phosphorylation triggers disruption of the EB3-SIAH-1 complex, resulting in EB3 stabilization during mitosis. Our results provide new insight into a regulatory mechanism of +TIPs in cell cycle transition.
Ban
Reiko
R
Department of Biochemistry, Shimane University School of Medicine, Izumo 693-8501; Department of Biochemistry, Nagoya University Graduate School of Medicine, Nagoya 466-8550. Electronic address: banrei@med.shimane-u.ac.jp.
Matsuzaki
Hideki
H
Institute for Enzyme Research, University of Tokushima, 3-15-18 Kuramoto, Tokushima 770-8503.
Akashi
Tomohiro
T
Division of Molecular Mycology and Medicine, Nagoya University Graduate School of Medicine, Nagoya 466-8550.
Sakashita
Gyosuke
G
Department of Biochemistry, Shimane University School of Medicine, Izumo 693-8501; Department of Biochemistry, Nagoya University Graduate School of Medicine, Nagoya 466-8550.
Taniguchi
Hisaaki
H
Institute for Enzyme Research, University of Tokushima, 3-15-18 Kuramoto, Tokushima 770-8503.
Park
Sam-Yong
SY
Protein Design Laboratory, Yokohama City University, Tsurumi, Yokohama 230-0045.
Tanaka
Hirofumi
H
School of Life Science, Tokyo University of Pharmacy and Life Science, Hachioji, Tokyo 192-0392, Japan.
Furukawa
Koichi
K
Department of Biochemistry, Nagoya University Graduate School of Medicine, Nagoya 466-8550.
Urano
Takeshi
T
Department of Biochemistry, Shimane University School of Medicine, Izumo 693-8501; Department of Biochemistry, Nagoya University Graduate School of Medicine, Nagoya 466-8550. Electronic address: turano@med.shimane-u.ac.jp.
D000595:Amino Acid Sequence; D000818:Animals; D064107:Aurora Kinase B; D064127:Aurora Kinases; D019556:COS Cells; D002522:Chlorocebus aethiops; D006367:HeLa Cells; D006801:Humans; D007527:Isoenzymes / Q000235:genetics / Q000378:metabolism; D008869:Microtubule-Associated Proteins / Q000235:genetics / Q000378:metabolism*; D008870:Microtubules / Q000378:metabolism*; D008938:Mitosis / Q000502:physiology*; D008969:Molecular Sequence Data; D009687:Nuclear Proteins / Q000235:genetics / Q000378:metabolism*; D017346:Protein Serine-Threonine Kinases / Q000235:genetics / Q000378:metabolism*; D034741:RNA, Small Interfering / Q000235:genetics / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D020798:Two-Hybrid System Techniques; D044767:Ubiquitin-Protein Ligases / Q000235:genetics / Q000378:metabolism*; D000098563:Seven in Absentia Proteins
D007527:Isoenzymes; C403664:MAPRE3 protein, human; D008869:Microtubule-Associated Proteins; D009687:Nuclear Proteins; D034741:RNA, Small Interfering; D011993:Recombinant Fusion Proteins; D044767:Ubiquitin-Protein Ligases; D000098563:Seven in Absentia Proteins; C576921:AURKB protein, human; D064107:Aurora Kinase B; D064127:Aurora Kinases; D017346:Protein Serine-Threonine Kinases
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