Identification of a novel substrate for TNFalpha-induced kinase NUAK2.
TNFalpha has multiple important cellular functions both in normal cells and in tumor cells. To explore the role of TNFalpha, we identified NUAK family, SNF1-like kinase 2 (NUAK2), as a TNFalpha-induced kinase by gene chip analysis. NUAK2 is known to be induced by various cellular stresses and involved in cell mortality, however, its substrate has never been identified. We developed original protocol of de novo screening for kinase substrates using an in vitro kinase assay and high performance liquid chromatography (HPLC). Using this procedure, we identified myosin phosphatase target subunit 1 (MYPT1) as a specific substrate for NUAK2. MYPT1 was phosphorylated at another site(s) by NUAK2, other than known Rho-kinase phosphorylation sites (Thr696 or Thr853) responsible for inhibition of myosin phosphatase activity. These data suggests different phosphorylation and regulation of MYPT1 activity by NUAK2.
Yamamoto
Hiroyuki
H
Department of Medical Information Science, Osaka University Graduate School of Medicine, Suita, Osaka 565-0871, Japan.
Takashima
Seiji
S
Shintani
Yasunori
Y
Yamazaki
Satoru
S
Seguchi
Osamu
O
Nakano
Atsushi
A
Higo
Shuichiro
S
Kato
Hisakazu
H
Liao
Yulin
Y
Asano
Yoshihiro
Y
Minamino
Tetsuo
T
Matsumura
Yasushi
Y
Takeda
Hiroshi
H
Kitakaze
Masafumi
M
D000595:Amino Acid Sequence; D002460:Cell Line; D006801:Humans; D008969:Molecular Sequence Data; D043263:Myosin-Light-Chain Phosphatase / Q000378:metabolism*; D010766:Phosphorylation; D017346:Protein Serine-Threonine Kinases / Q000235:genetics / Q000378:metabolism*; D012333:RNA, Messenger / Q000378:metabolism; D013379:Substrate Specificity; D013912:Threonine / Q000378:metabolism; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology / Q000502:physiology*
D012333:RNA, Messenger; D014409:Tumor Necrosis Factor-alpha; D013912:Threonine; C486061:NUAK2 protein, human; D017346:Protein Serine-Threonine Kinases; D043263:Myosin-Light-Chain Phosphatase; C490372:PPP1R12A protein, human