Proteomic and functional analysis of Argonaute-containing mRNA-protein complexes in human cells

被引：282
作者：
Hoeck, Julia
Weinmann, Lasse
Ender, Christine
Ruedel, Sabine
Kremmer, Elisabeth
Raabe, Monika
Urlaub, Henning
Meister, Gunter [1 ]
机构：
[1] Max Planck Inst Biochem, Lab RNA Biol, Martinsried, Germany
[2] GSF Munich, Inst Mol Immunol, Munich, Germany
[3] Max Planck Inst Biophys Chem, Bioanalyt Mass Spectrometry Grp, Gottingen, Germany
来源：
EMBO REPORTS | 2007年 / 8卷 / 11期
关键词：
Argonaute proteins; gene silencing; microRNA; RNA interference;
D O I：
10.1038/sj.embor.7401088
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：
Members of the Argonaute ( Ago) protein family associate with small RNAs and have important roles in RNA silencing. Here, we analysed Ago1- and Ago2- containing protein complexes in human cells. Separation of Ago- associated messenger ribonucleoproteins ( mRNPs) showed that Ago1 and Ago2 reside in three complexes with distinct Dicer and RNA- induced silencing complex activities. A comprehensive proteomic analysis of Ago- containing mRNPs identified a large number of proteins involved in RNA metabolism. By using co- immunoprecipitation experiments followed by RNase treatment, we biochemically mapped interactions within Ago mRNPs. Using reporter assays and knockdown experiments, we showed that the putative RNA- binding protein RBM4 is required for microRNA- guided gene regulation.
引用
收藏
页码：1052 / 1060
页数：9
相关论文
共 28 条
  • [1] 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
  • [2] Identification of human microRNA targets from isolated argonaute protein complexes
    Beitzinger, Michaela
    Peters, Lasse
    Zhu, Jia Yun
    Kremmer, Elisabeth
    Meister, Gunter
    [J]. RNA BIOLOGY, 2007, 4 (02) : 76 - 84
  • [3] Translation repression in human cells by microRNA-induced gene silencing requires RCK/p54
    Chu, Chia-ying
    Rana, Tariq M.
    [J]. PLOS BIOLOGY, 2006, 4 (07) : 1122 - 1136
  • [4] P-body formation is a consequence, not the cause, of RNA-mediated gene silencing
    Eulalio, Ana
    Behm-Ansmant, Isabelle
    Schweizer, Daniel
    Izaurralde, Elisa
    [J]. MOLECULAR AND CELLULAR BIOLOGY, 2007, 27 (11) : 3970 - 3981
  • [5] Post-transcriptional gene silencing by siRNAs and miRNAs
    Filipowicz, W
    Jaskiewicz, L
    Kolb, FA
    Pillai, RS
    [J]. CURRENT OPINION IN STRUCTURAL BIOLOGY, 2005, 15 (03) : 331 - 341
  • [6] FULALIO A, 2007, NAT REV MOL CELL BIO, V8, P9
  • [7] Gregory RI, 2005, CELL, V123, P631, DOI 10.1016/j.cell.2005.10.022
  • [8] The Microprocessor complex mediates the genesis of microRNAs
    Gregory, RI
    Yan, KP
    Amuthan, G
    Chendrimada, T
    Doratotaj, B
    Cooch, N
    Shiekhattar, R
    [J]. NATURE, 2004, 432 (7014) : 235 - 240
  • [9] Johnson SM, 2005, CELL, V120, P635, DOI 10.1016/j.cell.2005.01.014
  • [10] Identification of many microRNAs that copurify with polyribosomes in mammalian neurons
    Kim, J
    Krichevsky, A
    Grad, Y
    Hayes, GD
    Kosik, KS
    Church, GM
    Ruvkun, G
    [J]. PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 2004, 101 (01) : 360 - 365
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