title: Deep sequencing and proteomic analysis of the microRNA-induced silencing complex in human red blood cells.
abstract: During maturation, erythropoietic cells extrude their nuclei but retain their ability to respond to oxidant stress by tightly regulating protein translation. Several studies have reported microRNA-mediated regulation of translation during terminal stages of erythropoiesis, even after enucleation. In the present study, we performed a detailed examination of the endogenous microRNA machinery in human red blood cells using a combination of deep sequencing analysis of microRNAs and proteomic analysis of the microRNA-induced silencing complex. Among the 197 different microRNAs detected, miR-451a was the most abundant, representing more than 60% of all read sequences. In addition, miR-451a and its known target, 14-3-3ζ mRNA, were bound to the microRNA-induced silencing complex, implying their direct interaction in red blood cells. The proteomic characterization of endogenous Argonaute 2-associated microRNA-induced silencing complex revealed 26 cofactor candidates. Among these cofactors, we identified several RNA-binding proteins, as well as motor proteins and vesicular trafficking proteins. Our results demonstrate that red blood cells contain complex microRNA machinery, which might enable immature red blood cells to control protein translation independent of de novo nuclei information.
authors: Division of Hematology, University Children's Hospital, Zurich, Switzerland; Children's Research Center, University Children's Hospital, Zurich, Switzerland; Zurich Center for Integrative Human Physiology, University of Zurich, Zurich, Switzerland.
authors: Imane
authors: I
authors: Azzouzi
authors: Institute of Molecular Systems Biology, Eidgenössische Technische Hochschule (ETH), Zurich, Switzerland.
authors: Hansjoerg
authors: H
authors: Moest
authors: Institute of Molecular Systems Biology, Eidgenössische Technische Hochschule (ETH), Zurich, Switzerland.
authors: Bernd
authors: B
authors: Wollscheid
authors: Division of Hematology, University Children's Hospital, Zurich, Switzerland; Children's Research Center, University Children's Hospital, Zurich, Switzerland; Zurich Center for Integrative Human Physiology, University of Zurich, Zurich, Switzerland.
authors: Markus
authors: M
authors: Schmugge
authors: Division of Hematology, University Children's Hospital, Zurich, Switzerland; Children's Research Center, University Children's Hospital, Zurich, Switzerland. Electronic address: julia.eekels@kispi.uzh.ch.
authors: Julia J M
authors: JJM
authors: Eekels
authors: Division of Hematology, University Children's Hospital, Zurich, Switzerland; Children's Research Center, University Children's Hospital, Zurich, Switzerland.
authors: Oliver
authors: O
authors: Speer
mesh_terms: D048948:14-3-3 Proteins / Q000235:genetics / Q000378:metabolism; D060565:Argonaute Proteins / Q000235:genetics / Q000378:metabolism; D001483:Base Sequence; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D004912:Erythrocytes / Q000378:metabolism*; D059014:High-Throughput Nucleotide Sequencing / Q000379:methods*; D006801:Humans; D013058:Mass Spectrometry; D035683:MicroRNAs / Q000235:genetics* / Q000378:metabolism; D011485:Protein Binding; D020543:Proteome / Q000378:metabolism; D040901:Proteomics / Q000379:methods*; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D034743:RNA-Induced Silencing Complex / Q000235:genetics* / Q000378:metabolism; D012156:Reticulocytes / Q000378:metabolism; D012689:Sequence Homology, Nucleic Acid
chemical_list: D048948:14-3-3 Proteins; C466422:AGO2 protein, human; D060565:Argonaute Proteins; C531201:MIRN451 microRNA, human; D035683:MicroRNAs; D020543:Proteome; D012333:RNA, Messenger; D034743:RNA-Induced Silencing Complex