Identification of miRNA targets with stable isotope labeling by amino acids in cell culture.
miRNAs are small noncoding RNAs that regulate gene expression. We have used stable isotope labeling by amino acids in cell culture (SILAC) to investigate the effect of miRNA-1 on the HeLa cell proteome. Expression of 12 out of 504 investigated proteins was repressed by miRNA-1 transfection. This repressed set of genes significantly overlaps with miRNA-1 regulated genes that have been identified with DNA array technology and are predicted by computational methods. Moreover, we find that the 3'-untranslated region for the repressed set are enriched in miRNA-1 complementary sites. Our findings demonstrate that SILAC can be used for miRNA target identification and that one highly expressed miRNA can regulate the levels of many different proteins.
Vinther
Jeppe
J
Molecular Evolution Group, Institute of Molecular Biology and Physiology, University of Copenhagen, Universitetsparken 15, Building 10 DK-2100 Copenhagen Ø, Denmark. jvinther@bi.ku.dk
Hedegaard
Mads M
MM
Gardner
Paul P
PP
Andersen
Jens S
JS
Arctander
Peter
P
D020413:3' Untranslated Regions / Q000737:chemistry*; D000596:Amino Acids / Q000737:chemistry; D002247:Carbon Isotopes; D005786:Gene Expression Regulation; D006367:HeLa Cells; D006801:Humans; D013058:Mass Spectrometry; D035683:MicroRNAs / Q000502:physiology*; D009587:Nitrogen Isotopes; D020543:Proteome / Q000737:chemistry / Q000235:genetics*; D040901:Proteomics / Q000379:methods*
D020413:3' Untranslated Regions; D000596:Amino Acids; D002247:Carbon Isotopes; D035683:MicroRNAs; D009587:Nitrogen Isotopes; D020543:Proteome