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      mRNA silencing in erythroid differentiation: hnRNP K and hnRNP E1 regulate 15-lipoxygenase translation from the 3' end.

      Cell
      Amino Acid Sequence, Animals, Arachidonate 15-Lipoxygenase, genetics, Base Sequence, Cell Differentiation, Cell-Free System, Cytoplasm, metabolism, Erythrocytes, cytology, Erythropoiesis, HeLa Cells, Heterogeneous-Nuclear Ribonucleoprotein K, Heterogeneous-Nuclear Ribonucleoproteins, Humans, In Vitro Techniques, Molecular Sequence Data, Oligodeoxyribonucleotides, Protein Biosynthesis, RNA, Messenger, Rabbits, Reticulocytes, Ribonucleoproteins, Transfection

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          Abstract

          Although LOX mRNA accumulates early during differentiation, a differentiation control element in its 3' untranslated region confers translational silencing until late stage erythropoiesis. We have purified two proteins from rabbit reticulocytes that specifically mediate LOX silencing and identified them as hnRNPs K and E1. Transfection of hnRNP K and hnRNP E1 into HeLa cells specifically silenced the translation of reporter mRNAs bearing a differentiation control element in their 3' untranslated region. Silenced LOX mRNA in rabbit reticulocytes specifically coimmunoprecipitated with hnRNP K. In a reconstituted cell-free translation system, addition of recombinant hnRNP K and hnRNP E1 recapitulates this regulation via a specific inhibition of 80S ribosome assembly on LOX mRNA. Both proteins can control cap-dependent and internal ribosome entry site-mediated translation by binding to differentiation control elements. Our data suggest a specific cytoplasmic function for hnRNPs as translational regulatory proteins.

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