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Nucleolin binds to a subset of selenoprotein mRNAs and regulates their expression

Selenium, an essential trace element, is incorporated into selenoproteins as selenocysteine (Sec), the 21st amino acid. In order to synthesize selenoproteins, a translational reprogramming event must occur since Sec is encoded by the UGA stop codon. In mammals, the recoding of UGA as Sec depends on...

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Autores principales: Miniard, Angela C., Middleton, Lisa M., Budiman, Michael E., Gerber, Carri A., Driscoll, Donna M.
Formato: Texto
Lenguaje:English
Publicado: Oxford University Press 2010
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2919729/
https://www.ncbi.nlm.nih.gov/pubmed/20385601
http://dx.doi.org/10.1093/nar/gkq247
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author Miniard, Angela C.
Middleton, Lisa M.
Budiman, Michael E.
Gerber, Carri A.
Driscoll, Donna M.
author_facet Miniard, Angela C.
Middleton, Lisa M.
Budiman, Michael E.
Gerber, Carri A.
Driscoll, Donna M.
author_sort Miniard, Angela C.
collection PubMed
description Selenium, an essential trace element, is incorporated into selenoproteins as selenocysteine (Sec), the 21st amino acid. In order to synthesize selenoproteins, a translational reprogramming event must occur since Sec is encoded by the UGA stop codon. In mammals, the recoding of UGA as Sec depends on the selenocysteine insertion sequence (SECIS) element, a stem-loop structure in the 3′ untranslated region of the transcript. The SECIS acts as a platform for RNA-binding proteins, which mediate or regulate the recoding mechanism. Using UV crosslinking, we identified a 110 kDa protein, which binds with high affinity to SECIS elements from a subset of selenoprotein mRNAs. The crosslinking activity was purified by RNA affinity chromatography and identified as nucleolin by mass spectrometry analysis. In vitro binding assays showed that purified nucleolin discriminates among SECIS elements in the absence of other factors. Based on siRNA experiments, nucleolin is required for the optimal expression of certain selenoproteins. There was a good correlation between the affinity of nucleolin for a SECIS and its effect on selenoprotein expression. As selenoprotein transcript levels and localization did not change in siRNA-treated cells, our results suggest that nucleolin selectively enhances the expression of a subset of selenoproteins at the translational level.
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spelling pubmed-29197292010-08-11 Nucleolin binds to a subset of selenoprotein mRNAs and regulates their expression Miniard, Angela C. Middleton, Lisa M. Budiman, Michael E. Gerber, Carri A. Driscoll, Donna M. Nucleic Acids Res Molecular Biology Selenium, an essential trace element, is incorporated into selenoproteins as selenocysteine (Sec), the 21st amino acid. In order to synthesize selenoproteins, a translational reprogramming event must occur since Sec is encoded by the UGA stop codon. In mammals, the recoding of UGA as Sec depends on the selenocysteine insertion sequence (SECIS) element, a stem-loop structure in the 3′ untranslated region of the transcript. The SECIS acts as a platform for RNA-binding proteins, which mediate or regulate the recoding mechanism. Using UV crosslinking, we identified a 110 kDa protein, which binds with high affinity to SECIS elements from a subset of selenoprotein mRNAs. The crosslinking activity was purified by RNA affinity chromatography and identified as nucleolin by mass spectrometry analysis. In vitro binding assays showed that purified nucleolin discriminates among SECIS elements in the absence of other factors. Based on siRNA experiments, nucleolin is required for the optimal expression of certain selenoproteins. There was a good correlation between the affinity of nucleolin for a SECIS and its effect on selenoprotein expression. As selenoprotein transcript levels and localization did not change in siRNA-treated cells, our results suggest that nucleolin selectively enhances the expression of a subset of selenoproteins at the translational level. Oxford University Press 2010-08 2010-04-12 /pmc/articles/PMC2919729/ /pubmed/20385601 http://dx.doi.org/10.1093/nar/gkq247 Text en © The Author(s) 2010. Published by Oxford University Press. http://creativecommons.org/licenses/by-nc/2.5 This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/2.5), which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Molecular Biology
Miniard, Angela C.
Middleton, Lisa M.
Budiman, Michael E.
Gerber, Carri A.
Driscoll, Donna M.
Nucleolin binds to a subset of selenoprotein mRNAs and regulates their expression
title Nucleolin binds to a subset of selenoprotein mRNAs and regulates their expression
title_full Nucleolin binds to a subset of selenoprotein mRNAs and regulates their expression
title_fullStr Nucleolin binds to a subset of selenoprotein mRNAs and regulates their expression
title_full_unstemmed Nucleolin binds to a subset of selenoprotein mRNAs and regulates their expression
title_short Nucleolin binds to a subset of selenoprotein mRNAs and regulates their expression
title_sort nucleolin binds to a subset of selenoprotein mrnas and regulates their expression
topic Molecular Biology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2919729/
https://www.ncbi.nlm.nih.gov/pubmed/20385601
http://dx.doi.org/10.1093/nar/gkq247
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