Translation of 15-lipoxygenase mRNA is inhibited by a protein that binds to a repeated sequence in the 3' untranslated region.
During red blood cell differentiation, the mRNA encoding rabbit erythroid 15-lipoxygenase (LOX) is synthesized in the early stages of erythropoiesis, but is only activated for translation in peripheral reticulocytes. Erythroid LOX, which like other lipoxygenases catalyses the degradation of lipids, is unique in its ability to attack intact phospholipids and is the main factor responsible for the degradation of mitochondria during reticulocyte maturation. Strikingly, rabbit erythroid LOX mRNA has 10 tandem repeats of a slightly varied, pyrimidine-rich 19 nt motif in its 3'-untranslated region (3'-UTR). In this study we demonstrate, using gel retardation and UV-crosslinking assays, that this 3'-UTR segment specifically binds a 48 kDa reticulocyte protein. Furthermore, the interaction between the 3'-UTR LOX repeat motif and the 48 kDa protein, purified to homogeneity by specific RNA chromatography, is shown to be necessary and sufficient for specific translational repression of LOX as well as reporter mRNAs in vitro. To our knowledge this is the first case in which translation, presumably at the initiation step, is regulated by a defined protein-RNA interaction in the 3'-UTR.
Ostareck-Lederer
A
A
Institute of Biochemistry, Faculty of Medicine (Charité), Humboldt-University of Berlin, Germany.
Ostareck
D H
DH
Standart
N
N
Thiele
B J
BJ
D000818:Animals; D001093:Arachidonate 15-Lipoxygenase / Q000235:genetics*; D001483:Base Sequence; D001665:Binding Sites; D001798:Blood Proteins / Q000302:isolation & purification / Q000378:metabolism*; D002846:Chromatography, Affinity; D015536:Down-Regulation; D007438:Introns; D008969:Molecular Sequence Data; D009838:Oligodeoxyribonucleotides; D014176:Protein Biosynthesis; D012333:RNA, Messenger / Q000037:antagonists & inhibitors / Q000235:genetics* / Q000378:metabolism; D016601:RNA-Binding Proteins / Q000302:isolation & purification / Q000378:metabolism*; D011817:Rabbits; D012091:Repetitive Sequences, Nucleic Acid; D012156:Reticulocytes / Q000378:metabolism
D001798:Blood Proteins; D009838:Oligodeoxyribonucleotides; D012333:RNA, Messenger; D016601:RNA-Binding Proteins; D001093:Arachidonate 15-Lipoxygenase