Quantitative proteomics identify novel miR-155 target proteins

• Published in: PloS one Vol. 6; no. 7; p. e22146
• Main Authors: Lössner, Christopher, Meier, Jan, Warnken, Uwe, Rogers, Michael A, Lichter, Peter, Pscherer, Armin, Schnölzer, Martina
• Format: Journal Article
• Language: English
• Published: United States Public Library of Science 25.07.2011; Public Library of Science (PLoS)
• Subjects: Amino Acid Sequence; Amino acids; Amino Acids - chemistry; Amino Acids - metabolism; Annotations; Biology; Blotting, Western; Cancer; Cell culture; Cell cycle; Cloning; Deregulation; Down-Regulation - genetics; Gene expression; Growth factors; HEK293 Cells; HeLa Cells; Humans; Isotope Labeling; Labeling; Labelling; Luciferase; Medical research; MicroRNA; MicroRNAs; MicroRNAs - genetics; MicroRNAs - metabolism; miRNA; Molecular Sequence Annotation; Molecular Sequence Data; Nucleotides; Proteins; Proteins - genetics; Proteins - metabolism; Proteomics; Proteomics - methods; Reverse Transcriptase Polymerase Chain Reaction; Ribonucleic acid; RNA; Target recognition; Tissues; Transcription; Transcription (Genetics); United States > US; Germany; Singapore
• ISSN: 1932-6203; 1932-6203
• DOI: 10.1371/journal.pone.0022146

MicroRNAs are 22 nucleotides long non-coding RNAs and exert their function either by transcriptional or translational inhibition. Although many microRNA profiles in different tissues and disease states have already been discovered, only little is known about their target proteins. The microRNA miR-155 is deregulated in many diseases, including cancer, where it might function as an oncoMir. We employed a proteomics technique called "stable isotope labelling by amino acids in cell culture" (SILAC) allowing relative quantification to reliably identify target proteins of miR-155. Using SILAC, we identified 46 putative miR-155 target proteins, some of which were previously reported. With luciferase reporter assays, CKAP5 was confirmed as a new target of miR-155. Functional annotation of miR-155 target proteins pointed to a role in cell cycle regulation. To the best of our knowledge we have investigated for the first time miR-155 target proteins in the HEK293T cell line in large scale. In addition, by comparing our results to previously identified miR-155 target proteins in other cell lines, we provided further evidence for the cell line specificity of microRNAs.