Phosphorylation of human Argonaute proteins affects small RNA binding

被引:132
作者
Ruedel, Sabine [1 ,2 ]
Wang, Yanli [3 ]
Lenobel, Rene [4 ]
Koerner, Roman [4 ]
Hsiao, He-Hsuan [5 ]
Urlaub, Henning [5 ]
Patel, Dinshaw [3 ]
Meister, Gunter [1 ,2 ]
机构
[1] Max Planck Inst Biochem, CIPSM, Lab RNA Biol, D-82152 Martinsried, Germany
[2] Univ Regensburg, Dept Biochem, Lab RNA Biol, D-93053 Regensburg, Germany
[3] Mem Sloan Kettering Canc Ctr, Struct Biol Program, New York, NY 10065 USA
[4] Max Planck Inst Biochem, Dept Cell Biol, D-82152 Martinsried, Germany
[5] Max Planck Inst Biophys Chem, Bioanalyt Mass Spectrometry Grp, D-37077 Gottingen, Germany
关键词
AEOLICUS ARGONAUTE; SILENCING COMPLEX; CRYSTAL-STRUCTURE; MESSENGER-RNAS; MICRORNA; MIRNAS; MECHANISMS; LOCALIZATION; RECOGNITION; INHIBITION;
D O I
10.1093/nar/gkq1032
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Argonaute (Ago) proteins are highly conserved between species and constitute a direct-binding platform for small RNAs including short-interfering RNAs (siRNAs), microRNAs (miRNAs) and Piwi interacting RNAs (piRNAs). Small RNAs function as guides whereas Ago proteins are the actual mediators of gene silencing. Although the major steps in RNA-guided gene silencing have been elucidated, not much is known about Ago-protein regulation. Here we report a comprehensive analysis of Ago2 phosphorylation in human cells. We find that the highly conserved tyrosine Y529, located in the small RNA 5'-end-binding pocket of Ago proteins can be phosphorylated. By substituting Y529 with a negatively charged glutamate (E) mimicking a phosphorylated tyrosine, we show that small RNA binding is strongly reduced. Our data suggest that a negatively charged phospho-tyrosine generates a repulsive force that prevents efficient binding of the negatively charged 5' phosphate of the small RNA.
引用
收藏
页码:2330 / 2343
页数:14
相关论文
共 43 条
  • [1] Argonaute-2 Expression Is Regulated by Epidermal Growth Factor Receptor and Mitogen-Activated Protein Kinase Signaling and Correlates with a Transformed Phenotype in Breast Cancer Cells
    Adams, Brian D.
    Claffey, Kevin P.
    White, Bruce A.
    [J]. ENDOCRINOLOGY, 2009, 150 (01) : 14 - 23
  • [2] MRNA degradation by miRNAs and GW182 requires both CCR4:NOT deadenylase and DCP1:DCP2 decapping complexes
    Behm-Ansmant, Isabelle
    Rehwinkel, Jan
    Doerks, Tobias
    Stark, Alexander
    Bork, Peer
    Izaurralde, Elisa
    [J]. GENES & DEVELOPMENT, 2006, 20 (14) : 1885 - 1898
  • [3] Origins and Mechanisms of miRNAs and siRNAs
    Carthew, Richard W.
    Sontheimer, Erik J.
    [J]. CELL, 2009, 136 (04) : 642 - 655
  • [4] Strand-specific 5′-O-methylation of siRNA duplexes controls guide strand selection and targeting specificity
    Chen, Po Yu
    Weinmann, Lasse
    Gaidatzis, Dimos
    Pei, Yi
    Zavolan, Mihaela
    Tuschl, Thomas
    Meister, Gunter
    [J]. RNA, 2008, 14 (02) : 263 - 274
  • [5] A Human snoRNA with MicroRNA-Like Functions
    Ender, Christine
    Krek, Azra
    Friedlaender, Marc R.
    Beitzinger, Michaela
    Weinmann, Lasse
    Chen, Wei
    Pfeffer, Sebastien
    Rajewsky, Nikolaus
    Meister, Gunter
    [J]. MOLECULAR CELL, 2008, 32 (04) : 519 - 528
  • [6] Getting to the root of miRNA-Mediated gene silencing
    Eulalio, Ana
    Huntzinger, Eric
    Izaurralde, Elisa
    [J]. CELL, 2008, 132 (01) : 9 - 14
  • [7] P bodies: at the crossroads of post-transcriptional pathways
    Eulalio, Ana
    Behm-Ansmant, Isabelle
    Izaurralde, Elisa
    [J]. NATURE REVIEWS MOLECULAR CELL BIOLOGY, 2007, 8 (01) : 9 - 22
  • [8] A phosphorylated cytoplasmic autoantigen, GW182, associates with a unique population of human mRNAs within novel cytoplasmic speckles
    Eystathioy, T
    Chan, EKL
    Tenenbaum, SA
    Keene, JD
    Griffith, K
    Fritzler, MJ
    [J]. MOLECULAR BIOLOGY OF THE CELL, 2002, 13 (04) : 1338 - 1351
  • [9] Mammalian miRNA RISC Recruits CAF1 and PABP to Affect PABP-Dependent Deadenylation
    Fabian, Marc R.
    Mathonnet, Geraldine
    Sundermeier, Thomas
    Mathys, Hansruedi
    Zipprich, Jakob T.
    Svitkin, Yuri V.
    Rivas, Fabiola
    Jinek, Martin
    Wohischlegel, James
    Doudna, Jennifer A.
    Chen, Chyi-Ying A.
    Shyu, Ann-Bin
    Yates, John R., III
    Hannon, Gregory J.
    Filipowicz, Witold
    Duchaine, Thomas F.
    Sonenberg, Nahum
    [J]. MOLECULAR CELL, 2009, 35 (06) : 868 - 880
  • [10] Mechanisms of post-transcriptional regulation by microRNAs: are the answers in sight?
    Filipowicz, Witold
    Bhattacharyya, Suvendra N.
    Sonenberg, Nahum
    [J]. NATURE REVIEWS GENETICS, 2008, 9 (02) : 102 - 114