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Fine mapping of interactions between eEF1 alpha protein and 3 ' UTR of metallothionein-1 mRNA

Lookup NU author(s): Professor Zofia Chrzanowska-LightowlersORCiD, Professor John Hesketh

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Abstract

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 (eEF1 alpha) 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 eEF1 alpha, 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 eEF1 alpha and histidine-tRNA ligase, and that binding of eEF1 alpha is facilitated through domains I and III. (C) 2009 Elsevier Inc. All rights reserved.

Publication metadata

Author(s): Fan KB, Chrzanowska-Lightowlers ZMA, Hesketh JE

Publication type: Article

Publication status: Published

Journal: Biochemical and Biophysical Research Communications

Year: 2009

Volume: 386

Issue: 1

Pages: 82-88

Date deposited: 27/05/2010

ISSN (print): 0006-291X

ISSN (electronic): 1090-2104

Publisher: Academic Press

URL: http://dx.doi.org/10.1016/j.bbrc.2009.05.146

DOI: 10.1016/j.bbrc.2009.05.146

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