Abstract
Modulating the efficiency of translation plays an important role in a wide variety of cellular processes and is often mediated by trans-acting factors that interact with cis-acting sequences within the mRNA. Here we show that a cis-acting element, the Hsp83 degradation element (HDE), within the 3'-untranslated region of the Drosophila Hsp83 mRNA functions as a translational enhancer. We show that this element is bound by a multiprotein complex, and we identify components using a novel affinity-based method called tandem RNA affinity purification tagging. Three proteins (DDP1, Hrp48, and poly(A)-binding protein) are components of the HDE-binding complex and function in translational enhancement. Our data support a model whereby the HDE is composed of several cis-acting subelements that represent binding sites for trans-acting factors, and the combined action of these trans-acting factors underlies the ability of the HDE to stimulate translation.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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3' Untranslated Regions / genetics
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3' Untranslated Regions / metabolism*
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Animals
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Binding Sites / genetics
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DNA-Binding Proteins / genetics
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DNA-Binding Proteins / metabolism*
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Drosophila Proteins / biosynthesis
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Drosophila Proteins / genetics
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Drosophila Proteins / metabolism*
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Drosophila melanogaster
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Heat-Shock Proteins / biosynthesis
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Heat-Shock Proteins / genetics
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Heterogeneous-Nuclear Ribonucleoproteins / genetics
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Heterogeneous-Nuclear Ribonucleoproteins / metabolism*
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Multiprotein Complexes / genetics
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Multiprotein Complexes / metabolism*
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Poly(A)-Binding Proteins / genetics
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Poly(A)-Binding Proteins / metabolism*
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Protein Biosynthesis / physiology*
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RNA Stability / physiology
Substances
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3' Untranslated Regions
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DNA-Binding Proteins
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Dp1 protein, Drosophila
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Drosophila Proteins
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Heat-Shock Proteins
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Heterogeneous-Nuclear Ribonucleoproteins
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Hrb27C protein, Drosophila
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Hsp83 protein, Drosophila
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Multiprotein Complexes
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Poly(A)-Binding Proteins