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标题:The nuclear protein Artemis promotes AMPK activation by stabilizing the LKB1–AMPK complex
时间:2019-11-19 21:30:35
DOI:10.1016/j.bbrc.2012.09.140
作者:Koji Nakagawa; Yasuko Uehata; Mitsuteru Natsuizaka
关键词:AMPK;LKB1;Artemis;Signal transduction
出版源: Biochemical and biophysical research communicat... ,427 (4)
摘要:AMP-activated protein kinase (AMPK) is a hetero-trimeric Ser/Thr kinase composed of a catalytic alpha subunit and regulatory beta and gamma subunits; it functions as an energy sensor that controls cellular energy homeostasis. In response to an increased cellular AMP/ATP ratio, AMPK is activated by phosphorylation at Thr172 in the alpha-subunit by upstream AMPK kinases (AMPKKs), including tumor suppressor liver kinase B1 (LKB1). To elucidate more precise molecular mechanisms of AMPK activation, we performed yeast two-hybrid screening and isolated the complementary DNA (cDNA) encoding the nuclear protein Artemis/DNA cross-link repair 1C (DCLRE1C) as an AMPKalpha2-binding protein. Artemis was found to co-immunoprecipitate with AMPKalpha2, and the co-localization of Artemis with AMPKalpha2 in the nucleus was confirmed by immunofluorescence staining in U2OS cells. Moreover, over-expression of Artemis enhanced the phosphorylation of AMPKalpha2 and the AMPK substrate acetyl-CoA carboxylase (ACC). Conversely, RNAi-mediated knockdown of Artemis reduced AMPK and ACC phosphorylation. In addition, Artemis markedly increased the physical association between AMPKalpha2 and LKB1. Taken together, these results suggest that Artemis functions as a positive regulator of AMPK signaling by stabilizing the LKB1-AMPK complex. Copyright 2012 Elsevier Inc. All rights reserved.
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目录:
  • The nuclear protein Artemis promotes AMPK activation by stabilizing the LKB1–AMPK complex
    • 1 Introduction
    • 2 Materials and methods
      • 2.1 Cell culture and transfection
      • 2.2 Plasmid construction
      • 2.3 Yeast two-hybrid screening
      • 2.4 Immunoprecipitation, affinity pull-down and western blotting
      • 2.5 RNA interference
      • 2.6 Quantitative RT-PCR
      • 2.7 Indirect immunofluorescence
    • 3 Results
      • 3.1 Artemis physically associates and co-localiz
      • 3.2 Artemis promotes activation of AMPK
      • 3.3 Artemis stabilizes the LKB1–AMPK complex
    • 4 Discussion
    • Acknowledgments
    • References

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