Identification of novel argonaute-associated proteins.
RNA silencing processes are guided by small RNAs known as siRNAs and microRNAs (miRNAs) . They reside in ribonucleoprotein complexes, which guide the cleavage of complementary mRNAs or affect stability and translation of partial complementary mRNAs . Argonaute (Ago) proteins are at the heart of silencing effector complexes and bind the single-stranded siRNA and miRNA . Our biochemical analysis revealed that Ago2 is present in a pre-miRNA processing complex that is able to transfer the miRNA into a target-mRNA cleaving complex. To gain insight into the function and composition of RNA silencing complexes, we purified Ago1- and Ago2-containing complexes from human cells. Several known Ago1- and/or Ago2-associated proteins including Dicer were identified, but also two novel factors, the putative RNA helicase MOV10, and the RNA recognition motif (RRM)-containing protein TNRC6B/KIAA1093. The new proteins localize, similar to Ago proteins, to mRNA-degrading cytoplasmic P bodies, and they are functionally required to mediate miRNA-guided mRNA cleavage.
Meister
Gunter
G
Laboratory of RNA Molecular Biology, The Rockefeller University, 1230 York Avenue, Box 186, New York, New York 10021, USA. meister@biochem.mpg.de
Landthaler
Markus
M
Peters
Lasse
L
Chen
Po Yu
PY
Urlaub
Henning
H
Lührmann
Reinhard
R
Tuschl
Thomas
T
D060565:Argonaute Proteins; D015152:Blotting, Northern; D015153:Blotting, Western; D002460:Cell Line; D015852:Eukaryotic Initiation Factor-2; D039642:Eukaryotic Initiation Factors / Q000378:metabolism; D005455:Fluorescent Antibody Technique; D006801:Humans; D047908:Intracellular Signaling Peptides and Proteins / Q000235:genetics* / Q000378:metabolism; D013058:Mass Spectrometry; D035683:MicroRNAs / Q000378:metabolism; D009841:Oligonucleotides; D010448:Peptide Initiation Factors / Q000378:metabolism; D020365:RNA Helicases / Q000235:genetics* / Q000378:metabolism; D012333:RNA, Messenger / Q000378:metabolism*; D034741:RNA, Small Interfering / Q000235:genetics; D034743:RNA-Induced Silencing Complex / Q000235:genetics / Q000378:metabolism*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D043244:Ribonuclease III / Q000378:metabolism
C115848:AGO1 protein, human; C466422:AGO2 protein, human; D060565:Argonaute Proteins; D015852:Eukaryotic Initiation Factor-2; D039642:Eukaryotic Initiation Factors; D047908:Intracellular Signaling Peptides and Proteins; D035683:MicroRNAs; D009841:Oligonucleotides; D010448:Peptide Initiation Factors; D012333:RNA, Messenger; D034741:RNA, Small Interfering; D034743:RNA-Induced Silencing Complex; D043244:Ribonuclease III; D020365:RNA Helicases