title: Identification and characterization of a cell division-regulating kinase AKB1 (associated kinase of Trypanosoma brucei 14-3-3) through proteomics study of the Tb14-3-3 binding proteins.
abstract: We used a proteomics approach to identify the binding partners of Trypanosoma brucei 14-3-3 (Tb14-3-3) which led to the identification of a novel kinase, AKB1. The binding between these two proteins was mediated by an amphipathic groove structure in Tb14-3-3 and 1-438 amino acid sequence of AKB1. Recombinant AKB1 but not its ATP-binding-deficient mutant (DFG to NFG) possessed an auto-phosphorylation activity as well as a kinase activity towards a peptide substrate in vitro. However, the autophosphorylation was not required for the binding of AKB1 to Tb14-3-3. Interestingly, the kinase activity of AKB1 was inhibited by calcium, and the kinase was found to utilize GTP, and dATP in addition to ATP as phospho-donors. AKB1 formed homodimers through a leucine-zipper structure. Either knockdown of AKB1 or overexpression of AKB1, but not kinase-dead AKB1 mutant, deregulated cytokinesis and cell division, suggesting that kinase activity of AKB1 is crucial for its function. Furthermore, we showed that AKB1 exists in a detergent insoluble fraction. Laser confocal microscopy revealed that the majority of AKB1 is co-localized with α-tubulin. Taken together, these findings suggest that AKB1 might regulate cytokinesis and cell division by phosphorylating cytoskeleton-associated proteins.
authors: Division of Eukaryotic Microbiology, Department of Infectious Medicine, Kurume University School of Medicine, 67 Asahi-machi Kurume, Fukuoka 830-0011, Japan; Department of Virology, Institute of Tropical Medicine, Nagasaki University, 1-12-4 Sakamoto, Nagasaki 852-8523, Japan; Department of Protozoology, Institute of Tropical Medicine, Nagasaki University, 1-12-4 Sakamoto, Nagasaki 852-8523, Japan; Fukuoka Tokushukai Medical Center, 4-5 Sugu-Kita, Kasuga, Fukuoka 816-0864, Japan; Laboratory of Hygienic Chemistry, Daiichi University of Pharmacy, 22-1, Tamagawa-cho, Minami-ku, Fukuoka 815-8511, Japan; Department of Oncological Sciences, Experimental Therapeutics Institute, Mount Sinai School of Medicine, 1425 Madison Avenue, Room 1520, New York, NY 10029, USA; and Department of Radiation Oncology, Houston Methodist Research Institute, Weill Cornell Medical College, 6550 Fannin St, SM8-024, Houston, TX 77030, USA inouedna@med.kurume-u.ac.jp.
authors: Masahiro
authors: M
authors: Inoue
authors: Division of Eukaryotic Microbiology, Department of Infectious Medicine, Kurume University School of Medicine, 67 Asahi-machi Kurume, Fukuoka 830-0011, Japan; Department of Virology, Institute of Tropical Medicine, Nagasaki University, 1-12-4 Sakamoto, Nagasaki 852-8523, Japan; Department of Protozoology, Institute of Tropical Medicine, Nagasaki University, 1-12-4 Sakamoto, Nagasaki 852-8523, Japan; Fukuoka Tokushukai Medical Center, 4-5 Sugu-Kita, Kasuga, Fukuoka 816-0864, Japan; Laboratory of Hygienic Chemistry, Daiichi University of Pharmacy, 22-1, Tamagawa-cho, Minami-ku, Fukuoka 815-8511, Japan; Department of Oncological Sciences, Experimental Therapeutics Institute, Mount Sinai School of Medicine, 1425 Madison Avenue, Room 1520, New York, NY 10029, USA; and Department of Radiation Oncology, Houston Methodist Research Institute, Weill Cornell Medical College, 6550 Fannin St, SM8-024, Houston, TX 77030, USA.
authors: Kenta
authors: K
authors: Okamoto
authors: Division of Eukaryotic Microbiology, Department of Infectious Medicine, Kurume University School of Medicine, 67 Asahi-machi Kurume, Fukuoka 830-0011, Japan; Department of Virology, Institute of Tropical Medicine, Nagasaki University, 1-12-4 Sakamoto, Nagasaki 852-8523, Japan; Department of Protozoology, Institute of Tropical Medicine, Nagasaki University, 1-12-4 Sakamoto, Nagasaki 852-8523, Japan; Fukuoka Tokushukai Medical Center, 4-5 Sugu-Kita, Kasuga, Fukuoka 816-0864, Japan; Laboratory of Hygienic Chemistry, Daiichi University of Pharmacy, 22-1, Tamagawa-cho, Minami-ku, Fukuoka 815-8511, Japan; Department of Oncological Sciences, Experimental Therapeutics Institute, Mount Sinai School of Medicine, 1425 Madison Avenue, Room 1520, New York, NY 10029, USA; and Department of Radiation Oncology, Houston Methodist Research Institute, Weill Cornell Medical College, 6550 Fannin St, SM8-024, Houston, TX 77030, USA.
authors: Haruki
authors: H
authors: Uemura
authors: Division of Eukaryotic Microbiology, Department of Infectious Medicine, Kurume University School of Medicine, 67 Asahi-machi Kurume, Fukuoka 830-0011, Japan; Department of Virology, Institute of Tropical Medicine, Nagasaki University, 1-12-4 Sakamoto, Nagasaki 852-8523, Japan; Department of Protozoology, Institute of Tropical Medicine, Nagasaki University, 1-12-4 Sakamoto, Nagasaki 852-8523, Japan; Fukuoka Tokushukai Medical Center, 4-5 Sugu-Kita, Kasuga, Fukuoka 816-0864, Japan; Laboratory of Hygienic Chemistry, Daiichi University of Pharmacy, 22-1, Tamagawa-cho, Minami-ku, Fukuoka 815-8511, Japan; Department of Oncological Sciences, Experimental Therapeutics Institute, Mount Sinai School of Medicine, 1425 Madison Avenue, Room 1520, New York, NY 10029, USA; and Department of Radiation Oncology, Houston Methodist Research Institute, Weill Cornell Medical College, 6550 Fannin St, SM8-024, Houston, TX 77030, USA.
authors: Kouichi
authors: K
authors: Yasuda
authors: Division of Eukaryotic Microbiology, Department of Infectious Medicine, Kurume University School of Medicine, 67 Asahi-machi Kurume, Fukuoka 830-0011, Japan; Department of Virology, Institute of Tropical Medicine, Nagasaki University, 1-12-4 Sakamoto, Nagasaki 852-8523, Japan; Department of Protozoology, Institute of Tropical Medicine, Nagasaki University, 1-12-4 Sakamoto, Nagasaki 852-8523, Japan; Fukuoka Tokushukai Medical Center, 4-5 Sugu-Kita, Kasuga, Fukuoka 816-0864, Japan; Laboratory of Hygienic Chemistry, Daiichi University of Pharmacy, 22-1, Tamagawa-cho, Minami-ku, Fukuoka 815-8511, Japan; Department of Oncological Sciences, Experimental Therapeutics Institute, Mount Sinai School of Medicine, 1425 Madison Avenue, Room 1520, New York, NY 10029, USA; and Department of Radiation Oncology, Houston Methodist Research Institute, Weill Cornell Medical College, 6550 Fannin St, SM8-024, Houston, TX 77030, USA.
authors: Yoshihiko
authors: Y
authors: Motohara
authors: Division of Eukaryotic Microbiology, Department of Infectious Medicine, Kurume University School of Medicine, 67 Asahi-machi Kurume, Fukuoka 830-0011, Japan; Department of Virology, Institute of Tropical Medicine, Nagasaki University, 1-12-4 Sakamoto, Nagasaki 852-8523, Japan; Department of Protozoology, Institute of Tropical Medicine, Nagasaki University, 1-12-4 Sakamoto, Nagasaki 852-8523, Japan; Fukuoka Tokushukai Medical Center, 4-5 Sugu-Kita, Kasuga, Fukuoka 816-0864, Japan; Laboratory of Hygienic Chemistry, Daiichi University of Pharmacy, 22-1, Tamagawa-cho, Minami-ku, Fukuoka 815-8511, Japan; Department of Oncological Sciences, Experimental Therapeutics Institute, Mount Sinai School of Medicine, 1425 Madison Avenue, Room 1520, New York, NY 10029, USA; and Department of Radiation Oncology, Houston Methodist Research Institute, Weill Cornell Medical College, 6550 Fannin St, SM8-024, Houston, TX 77030, USA.
authors: Kouichi
authors: K
authors: Morita
authors: Division of Eukaryotic Microbiology, Department of Infectious Medicine, Kurume University School of Medicine, 67 Asahi-machi Kurume, Fukuoka 830-0011, Japan; Department of Virology, Institute of Tropical Medicine, Nagasaki University, 1-12-4 Sakamoto, Nagasaki 852-8523, Japan; Department of Protozoology, Institute of Tropical Medicine, Nagasaki University, 1-12-4 Sakamoto, Nagasaki 852-8523, Japan; Fukuoka Tokushukai Medical Center, 4-5 Sugu-Kita, Kasuga, Fukuoka 816-0864, Japan; Laboratory of Hygienic Chemistry, Daiichi University of Pharmacy, 22-1, Tamagawa-cho, Minami-ku, Fukuoka 815-8511, Japan; Department of Oncological Sciences, Experimental Therapeutics Institute, Mount Sinai School of Medicine, 1425 Madison Avenue, Room 1520, New York, NY 10029, USA; and Department of Radiation Oncology, Houston Methodist Research Institute, Weill Cornell Medical College, 6550 Fannin St, SM8-024, Houston, TX 77030, USA.
authors: Makoto
authors: M
authors: Hiromura
authors: Division of Eukaryotic Microbiology, Department of Infectious Medicine, Kurume University School of Medicine, 67 Asahi-machi Kurume, Fukuoka 830-0011, Japan; Department of Virology, Institute of Tropical Medicine, Nagasaki University, 1-12-4 Sakamoto, Nagasaki 852-8523, Japan; Department of Protozoology, Institute of Tropical Medicine, Nagasaki University, 1-12-4 Sakamoto, Nagasaki 852-8523, Japan; Fukuoka Tokushukai Medical Center, 4-5 Sugu-Kita, Kasuga, Fukuoka 816-0864, Japan; Laboratory of Hygienic Chemistry, Daiichi University of Pharmacy, 22-1, Tamagawa-cho, Minami-ku, Fukuoka 815-8511, Japan; Department of Oncological Sciences, Experimental Therapeutics Institute, Mount Sinai School of Medicine, 1425 Madison Avenue, Room 1520, New York, NY 10029, USA; and Department of Radiation Oncology, Houston Methodist Research Institute, Weill Cornell Medical College, 6550 Fannin St, SM8-024, Houston, TX 77030, USA.
authors: E Premkumar
authors: EP
authors: Reddy
authors: Division of Eukaryotic Microbiology, Department of Infectious Medicine, Kurume University School of Medicine, 67 Asahi-machi Kurume, Fukuoka 830-0011, Japan; Department of Virology, Institute of Tropical Medicine, Nagasaki University, 1-12-4 Sakamoto, Nagasaki 852-8523, Japan; Department of Protozoology, Institute of Tropical Medicine, Nagasaki University, 1-12-4 Sakamoto, Nagasaki 852-8523, Japan; Fukuoka Tokushukai Medical Center, 4-5 Sugu-Kita, Kasuga, Fukuoka 816-0864, Japan; Laboratory of Hygienic Chemistry, Daiichi University of Pharmacy, 22-1, Tamagawa-cho, Minami-ku, Fukuoka 815-8511, Japan; Department of Oncological Sciences, Experimental Therapeutics Institute, Mount Sinai School of Medicine, 1425 Madison Avenue, Room 1520, New York, NY 10029, USA; and Department of Radiation Oncology, Houston Methodist Research Institute, Weill Cornell Medical College, 6550 Fannin St, SM8-024, Houston, TX 77030, USA.
authors: Toshihide
authors: T
authors: Fukuma
authors: Division of Eukaryotic Microbiology, Department of Infectious Medicine, Kurume University School of Medicine, 67 Asahi-machi Kurume, Fukuoka 830-0011, Japan; Department of Virology, Institute of Tropical Medicine, Nagasaki University, 1-12-4 Sakamoto, Nagasaki 852-8523, Japan; Department of Protozoology, Institute of Tropical Medicine, Nagasaki University, 1-12-4 Sakamoto, Nagasaki 852-8523, Japan; Fukuoka Tokushukai Medical Center, 4-5 Sugu-Kita, Kasuga, Fukuoka 816-0864, Japan; Laboratory of Hygienic Chemistry, Daiichi University of Pharmacy, 22-1, Tamagawa-cho, Minami-ku, Fukuoka 815-8511, Japan; Department of Oncological Sciences, Experimental Therapeutics Institute, Mount Sinai School of Medicine, 1425 Madison Avenue, Room 1520, New York, NY 10029, USA; and Department of Radiation Oncology, Houston Methodist Research Institute, Weill Cornell Medical College, 6550 Fannin St, SM8-024, Houston, TX 77030, USA.
authors: Nobuo
authors: N
authors: Horikoshi
mesh_terms: D048948:14-3-3 Proteins / Q000378:metabolism*; D002118:Calcium / Q000378:metabolism; D002455:Cell Division; D011494:Protein Kinases / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D040901:Proteomics; D014346:Trypanosoma brucei brucei / Q000201:enzymology* / Q000378:metabolism
chemical_list: D048948:14-3-3 Proteins; D011494:Protein Kinases; D002118:Calcium
keywords: 14-3-3; Kinase; Proteomics; Trypanosoma brucei