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Fine mapping of interactions between eEF1α protein and 3′UTR of metallothionein-1 mRNA

The localization of metallothionein-1 (MT-1) mRNA to the perinuclear cytoskeleton is determined by a signal in the 3′untranslated region (3′UTR) and trans-acting binding proteins. The present study carried out detailed mapping of this signal and further characterized the binding to elongation factor...

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Detalles Bibliográficos
Autores principales: Fan, Kunbo, Chrzanowska-Lightowlers, Zofia M.A., Hesketh, John E.
Formato: Texto
Lenguaje:English
Publicado: Academic Press 2009
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2719692/
https://www.ncbi.nlm.nih.gov/pubmed/19501573
http://dx.doi.org/10.1016/j.bbrc.2009.05.146
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author Fan, Kunbo
Chrzanowska-Lightowlers, Zofia M.A.
Hesketh, John E.
author_facet Fan, Kunbo
Chrzanowska-Lightowlers, Zofia M.A.
Hesketh, John E.
author_sort Fan, Kunbo
collection PubMed
description The localization of metallothionein-1 (MT-1) mRNA to the perinuclear cytoskeleton is determined by a signal in the 3′untranslated region (3′UTR) and trans-acting binding proteins. The present study carried out detailed mapping of this signal and further characterized the binding to elongation factor 1 alpha (eEF1α) and other interacting proteins. Electrophoresis mobility shift assays demonstrated that shortening of a stem region proximal to nucleotides 66–76 abrogated binding. Full length recombinant rat eEF1α, and independently domains I and III, formed complexes with the mRNA. Proteins binding to biotinylated MT-1 3′UTR sequences were isolated using RNA-affinity techniques, and mass spectrometry identified histidine-tRNA ligase as one of the major MT-1 3′UTR binding proteins. We conclude that a 5-bp internal stem in the MT-1 3′UTR is critical for binding of eEF1α and histidine-tRNA ligase, and that binding of eEF1α is facilitated through domains I and III.
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spelling pubmed-27196922009-08-18 Fine mapping of interactions between eEF1α protein and 3′UTR of metallothionein-1 mRNA Fan, Kunbo Chrzanowska-Lightowlers, Zofia M.A. Hesketh, John E. Biochem Biophys Res Commun Article The localization of metallothionein-1 (MT-1) mRNA to the perinuclear cytoskeleton is determined by a signal in the 3′untranslated region (3′UTR) and trans-acting binding proteins. The present study carried out detailed mapping of this signal and further characterized the binding to elongation factor 1 alpha (eEF1α) and other interacting proteins. Electrophoresis mobility shift assays demonstrated that shortening of a stem region proximal to nucleotides 66–76 abrogated binding. Full length recombinant rat eEF1α, and independently domains I and III, formed complexes with the mRNA. Proteins binding to biotinylated MT-1 3′UTR sequences were isolated using RNA-affinity techniques, and mass spectrometry identified histidine-tRNA ligase as one of the major MT-1 3′UTR binding proteins. We conclude that a 5-bp internal stem in the MT-1 3′UTR is critical for binding of eEF1α and histidine-tRNA ligase, and that binding of eEF1α is facilitated through domains I and III. Academic Press 2009-08-14 /pmc/articles/PMC2719692/ /pubmed/19501573 http://dx.doi.org/10.1016/j.bbrc.2009.05.146 Text en © 2009 Elsevier Inc. https://creativecommons.org/licenses/by/3.0/ Open Access under CC BY 3.0 (https://creativecommons.org/licenses/by/3.0/) license
spellingShingle Article
Fan, Kunbo
Chrzanowska-Lightowlers, Zofia M.A.
Hesketh, John E.
Fine mapping of interactions between eEF1α protein and 3′UTR of metallothionein-1 mRNA
title Fine mapping of interactions between eEF1α protein and 3′UTR of metallothionein-1 mRNA
title_full Fine mapping of interactions between eEF1α protein and 3′UTR of metallothionein-1 mRNA
title_fullStr Fine mapping of interactions between eEF1α protein and 3′UTR of metallothionein-1 mRNA
title_full_unstemmed Fine mapping of interactions between eEF1α protein and 3′UTR of metallothionein-1 mRNA
title_short Fine mapping of interactions between eEF1α protein and 3′UTR of metallothionein-1 mRNA
title_sort fine mapping of interactions between eef1α protein and 3′utr of metallothionein-1 mrna
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2719692/
https://www.ncbi.nlm.nih.gov/pubmed/19501573
http://dx.doi.org/10.1016/j.bbrc.2009.05.146
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